Protocols   Select Another Resource Report Type

Authors: peijun he
Group: Coronavirus Method Development Community, XPRIZE Rapid Covid Testing, Highfield Diagnostics
Summary: Coronavirus Lateral Flow Assay (LFA) operation protocol

Proper citation: peijun he 2020. Coronavirus Lateral Flow Assay (LFA) operation protocol. protocols.io dx.doi.org/10.17504/protocols.io.bksjkwcn Copy   

•   Source: Protocols.io

Authors: Diego Lai
Group: Coronavirus Method Development Community, XPRIZE Rapid Covid Testing
Summary: LooK SPOT COVID-19 antigen rapid test is a lateral flow immunoassay intended for the qualitative detection of nucleocapsid protein antigen from SARS-CoV-2 in nasal swabs from patients suspected of COVID-19 within the first seven (7) days of symptom onset. Testing is limited to laboratories certified under the Clinical Laboratory Improvement Amendments of 1988 (CLIA), 42 U.S.C. §263a, that meet the requirements to perform moderate complexity tests. This test is authorized for use at the Point of Care (POC), i.e., in a patient care settings operating under a CLIA Certificate of Waiver, Certificate of Compliance, or Certificate of Accreditation.].Results are for the identification of SARS-CoV-2 nucleocapsid protein antigen. The antigen is generally detectable in nasal samples during the acute phase of infection. Positive results indicate the presence of viral antigens, but the clinical correlation with patient history and other diagnostic information is necessary to determine infection status. Positive results do not rule out a bacterial infection or co-infection with other viruses. The agent detected may not be the definite cause of thedisease. Laboratories within the United States and its territories are required to report all positive results to the appropriate public health authorities.Negative results should be treated as presumptive, and do not rule out SARS-CoV-2 infection, and should not be used as the sole basis for treatment or patient management decisions, including infection control decisions. Negative results should be considered in the context of a patient's recent exposures, history, and the presence of clinical signs and symptoms consistent with COVID-19, and confirmed with a molecular assay, if necessary, for patient management. LooK SPOT COVID-19 Antigen Rapid Test can deliver a diagnosis of the SARS-CoV-2 virus detection between 5 to 8 minutes by using machine learning AI. LooK SPOT’s AI algorithm has high accuracy and can identify the fluorescence response when human eyes cannot identify the low positive cases. Healthcare responders in the COVID-19 test sites often need to make time-sensitive decisions to determine the test results during the time many patients are within their vicinity. But the tempo, volume, stress, fatigue, lighting, fear, and various other factors can overwhelm healthcare responders when making the visual interpretation of antigen test results. It is of paramount importance to reduce healthcare responders' cognitive load by providing accurate test results in an easy-to-read format. LooK SPOT COVID-19 Antigen Rapid Test is a COVID-19 rapid test solution designed with a tactical edge to fight COVID-19. The LooK SPOT COVID-19 Antigen Rapid Test is intended for use at the Point of Care (POC) settings by trained personnel specifically instructed and trained in vitro diagnostic procedures. It is only for use under the Food and Drug Administration's Emergency Use Authorization.

Proper citation: Diego Lai 2020. LooK SPOT Antigen Rapid Test System. protocols.io dx.doi.org/10.17504/protocols.io.bp32mqqe Copy   

•   Source: Protocols.io

Authors: yliu
Group: OnsiteGene 1, XPRIZE Rapid Covid Testing
Summary: This OnsiteGene protocol is designed for testing the nasal swab samples without nucleic acid extraction. It uses the Star Array® Hi-SenseTM COVID-19 Molecular Testing Kit 1.0 in the one-step real-time RT-qPCR test assay to qualitatively detect RNA from SARS-CoV-2 virus in the human respiratory specimen. It combines the reverse transcription technology and real-time PCR method to provide accurate detection of the SARS-CoV-2 coronavirus. Along with the Star Array® SATM Direct Extract Buffer, the protocol supports direct amplification without the need of specific RNA extraction equipment and kit. It significantly reduces the loss of RNA from the sample extraction and purification process, saves time and workload from sample preparation, and minimized the burdens on supply chain. The triplex fluorescence design of the kit simultaneously detects the N1 and Orf1ab genes of the virus and the human RNase P gene as an internal control to ensure the sample quality. This protocol uses the Star Array® XDiveTM Superfast Real-Time RT-qPCR instrument to perform 40-cycle PCR in 8 minutes, and can test up to 16 samples or controls in each run. The total protocol time from sample collection to data interpretation is less than 11 minutes.

Proper citation: yliu 2020. OnsiteGene 1 Protocol Nasal Direct. protocols.io dx.doi.org/10.17504/protocols.io.bkudkws6 Copy   

•   Source: Protocols.io

Authors: Monica Simion, Alexandru Salceanu
Group: Coronavirus Method Development Community, covid19-test, XPRIZE Rapid Covid Testing

Proper citation: Monica Simion, Alexandru Salceanu 2020. microfluidic sars cov 2 lamp protocol. protocols.io dx.doi.org/10.17504/protocols.io.bkjjkukn Copy   

•   Source: Protocols.io

Authors: lisa.diamond
Group: XPRIZE Rapid Covid Testing
Summary: Pinpoint's nanosensor-based platform detects SARS-CoV-2 antigen by (1) measuring baseline current through our functionalized nanosensor chip in ordinary PBS buffer; (2) measuring the change in current at specific voltage ranges after insertion of the specimen as a result of binding between probe aptamer and target antigen; (3) computing delta between these two measurements. These measurements and computations are performed internally in quadruplicate, with four separately measured nanosensor channels. Consolidated results are reported to the user as {Detected; Not Detected; Inconclusive; or Error}.

Proper citation: lisa.diamond 2020. Pinpoint Science 30-Second Covid-19 Test Protocol. protocols.io dx.doi.org/10.17504/protocols.io.bkxmkxk6 Copy   

•   Source: Protocols.io

Authors: Lyle J. Arnold Ph.D., Stella M. Sung Ph.D.
Group: Coronavirus Method Development Community, XPRIZE Rapid Covid Testing
Summary: This protocol is for the Aegea Biotechnologies rapid PCR-based SARS-CoV-2 test. This assay uses patented "Switch-Blocker" technology as well as taqman probes to test for presence of SARS-CoV-2 and simultaneously orthogonally validate. A single amplification reaction is performed, and "Switch Blocker" is used on the forward strand and taqman is used on the reverse strand. The assay design has high sensitivity & specificity--single nucleotide level. Moreover, it is able to detect the SARS-CoV-2 L strain vs the SARS-CoV-2 S strain. The test can be adapted to point of care (Roche LIAT) as well as for different SARS-CoV-2 strains as the virus mutates. A next generation version of the assay could identify the presence of the SARS-CoV-2 L/S strains vs. influenza A/B. Finally, because of the sensitivity and specificity, the Aegea PCR-based SARS-CoV-2 test should be able to use saliva samples, and it is suitable for pooled testing. This protocol is designed for high throughput PCR (96 or 384 well plate formats).Keywords: PCR, COVID-19, coronavirus, SARS-CoV-2, high throughput, multiplex, Switch-Blocker, taqman, high sensitivity, high specificity, accurate, pooling, saliva, strain types, L-strain, S-strain, combination SARS-CoV-2 and influenza

Proper citation: Lyle J. Arnold Ph.D., Stella M. Sung Ph.D. 2020. Aegea Biotechnologies rapid PCR SARS-CoV-2 test (high sensitivity & specificity; able to detect different strain types). protocols.io dx.doi.org/10.17504/protocols.io.bk2dkya6 Copy   

•   Source: Protocols.io

Authors: Noah Toppings
Group: XPRIZE Rapid Covid Testing

Proper citation: Noah Toppings 2020. RT-LAMP Reaction. protocols.io dx.doi.org/10.17504/protocols.io.bkp7kvrn Copy   

•   Source: Protocols.io

Authors: Mario Thomas, Jasmine Sollen, Natalia Ivanova, Michelle Feng, Reda Fayek, Heidi Abdilla, Stephanie Lim, Amanda Naaum
Group: Coronavirus Method Development Community, XPRIZE Rapid Covid Testing, SANATA
Summary: This procedure outlines the protocol for testing for SARS-CoV-2 using a saliva sample collected from an individual. The purpose of this test is to detect low levels of SARS-CoV-2 antigen at a higher sensitivity. Precision Biomonitoring Inc. developed an ultra-rapid digital, disposable, highly-sensitive and inexpensive testing device used for screening purposes. The mobile app complementary to this medical device is connected through Bluetooth. Using this innovation, the user can be tested at point-of-care (POC) by a health care professional, and obtain qualitative results.

Proper citation: Mario Thomas, Jasmine Sollen, Natalia Ivanova, Michelle Feng, Reda Fayek, Heidi Abdilla, Stephanie Lim, Amanda Naaum 2020. XPRIZE SANATA Protocol for Saliva LFIA Test. protocols.io dx.doi.org/10.17504/protocols.io.bqfrmtm6 Copy   

•   Source: Protocols.io

Authors: Ruth Polachek, doreen
Group: XPRIZE Rapid Covid Testing
Summary: LessTests SARS-CoV2 general diagnosis protocol, using LessTests software solution.The LessTests method is agnostic to the labware, and has already been demonstrated as successful using multiple device and kit combinations.

Proper citation: Ruth Polachek, doreen 2020. LessTests. protocols.io dx.doi.org/10.17504/protocols.io.bkiakuae Copy   

•   Source: Protocols.io

Authors: John Daniels
Group: Coronavirus Method Development Community, Front Line Technologies, XPRIZE Rapid Covid Testing, Team Exhaled Breath Condensate
Summary: The Kinaptic mask-based COVID-19 testing system uses a unique Exhaled Breath Condensate (EBC) collector that converts breath vapor to a liquid biosample for rapid testing and wireless automatic contact tracing. Exhaled Breath Condensate (EBC) is collected from a mask-based EBC collector as a low cost disposable component. Our electronic biosensor detects even low concentrations of virus biomarkers if they are present in the EBC sample. Corona viruses have slowly evolving N-protein and quickly evolving S-protein. N-Protein testing screens for any corona virus infection / S-Protein screens for only COVID-19 infection. The Kinaptic mask-based COVID-19 testing system is a “two-stage filter”: 1) catch only of N-protein biomarker = trigger warning to get more stringent diagnostic and contact trace; 2) catch both N- and S-, or just S- biomarkers = more urgent need for immediate quarantine and medical attention. As the target biomarkers bind to the capture molecules, the electrical conductivity of the electronic biosensor changes. This change in conductive is amplified by our bluetooth electronics, analyzed to determine the presence and quantity of the target biomarker, and the test result wireless transmitted to a smartphone or directly to the cloud.

Proper citation: John Daniels 2020. Mask-Based Covid-10 testing system using Exhaled Breath Condensate. protocols.io dx.doi.org/10.17504/protocols.io.bkzzkx76 Copy   

•   Source: Protocols.io

Authors: Shripal Gandhi, Santo Purnama, Keyur Patel, Praveen Sukumara, Dr Rinu R Ravi
Group: Coronavirus Method Development Community, SensingSelfMission, XPRIZE Rapid Covid Testing
Summary: Summary:The S1 COVID-19 Rapid Antigen test kit developed by Sensing Self Pte. Ltd. can quickly and qualitatively detect the spike glycoprotein of novel coronavirus (SARS-COV-2) in human saliva/sputum/stool samples. It can be used as an aid for COVID-19 diagnosis.Objective:According to the clinical trial plan, the S1 COVID-19 Rapid Antigen test kit or “test reagent”, is to test saliva/sputum/stool samples from healthy subjects and confirmed COVID-19 patients. Test results are compared with another commercial SARS-COV-2 nucleic acid detection kit with CFDA approval, which is defined as the “gold standard”. The sensitivity, specificity, and total agreement rate are used to evaluate the feasibility of the test reagent in clinical applications.Method:A collection of clinical samples were examined by the S1 COVID-19 Rapid Antigen test kit and the gold standard SARS-COV-2 nucleic acid detection kit in parallel, to calculate the clinical sensitivity, clinical specificity and total agreement rate of the test reagent.Results:Compared to the gold standard, the clinical sensitivity of test reagent reached 90.0%, the clinical specificity reached 100.0%, and the total coincidence rate reached 95.0%.Conclusions:The performance of test reagent has a high agreement rate with the gold standard, proving its good feasibility in diagnosing suspected COVID-19 cases.

Proper citation: Shripal Gandhi, Santo Purnama, Keyur Patel, Praveen Sukumara, Dr Rinu R Ravi 2020. SensingSelf S1 Rapid Antigen Test (Saliva/Sputum/Stool). protocols.io dx.doi.org/10.17504/protocols.io.bj6ykrfw Copy   

•   Source: Protocols.io

Authors: steve.puts
Group: Coronavirus Method Development Community, Mmolecular, XPRIZE Rapid Covid Testing
Summary: The Direct One-Step RT-qPCR COVID-19 Test Kit is designed for quantitativereal-time analysis of target RNA directly from, swabs without the requirement of any priorRNA purification steps.Shipping: shipped on blue ice Storage Conditions: store at -20 °C Additional Storage Conditions: avoid freeze/thaw cycles stable at 4 °C for up to 4 weeks Shelf Life: 12 monthsForm: liquid / lyophilized

Proper citation: steve.puts 2020. Mmolecular COVID-19 Extraction-Free Direct-One-Step Fast Cycling Protocol. protocols.io dx.doi.org/10.17504/protocols.io.bkjpkumn Copy   

•   Source: Protocols.io

Authors: ayten
Group: XPRIZE Rapid Covid Testing

Proper citation: ayten 2020. Easy Covid LAMP. protocols.io dx.doi.org/10.17504/protocols.io.bk6gkzbw Copy   

•   Source: Protocols.io

Authors: Ilse Pretorius
Group: Electronic Pathogen Detection, XPRIZE Rapid Covid Testing
Summary: Electronic pathogen detection (EPD) is a non - invasive, rapid, affordable, point- of- care test, for Covid 19 resulting from infection with SARS-CoV-2 virus.EPD scanning technology, is a completely novel technique, designed to assess the human body for the presence or absence of a pathogen.The technology implements a unique scanning method, by delivering an extremely low electrical current to the skin, and then recording the resulting physiological response to this stimulation, from the skin.Although the technique is unique, there are similarities to existing technologies that can be used to explain the modus operandi, of this novel technology. The microcurrent delivery component of the diagnostic technology can be loosely described as containing elements analogous to Microcurrent Electrical Therapy (MET) and TENS (Trans dermal electrical nervous stimulation), whereas the response component of the technology can be seen as similar to Electrocardiography (ECG), Elelctro-enchephalogram (EEG) the Electrodermal Response (EDR), and Body Impedance Assessment (BIA).All of these modalities are widely used, considered safe and have been extensively researched.The skin offers convenient access to study the nervous system, not only is it richly innervated, but it also originates from the same embryonic layer as the nervous system. It is well known that the immune system and immune response is regulated by the neurological system.EPD technology uses this physiological platform offered by the skin to access real-time information of the immune response and compares this physiological information to electronically generated pathogen molecular patterns via a proprietary algorythm.The resulting dermal response , is recorded and displayed on a computer screen as a graph.The graph consists of peaks and troughs.This graph is then statistically verified and analyzed in a proprietary manner.The result of this analysis is then compared to the pathogen dataset that is pre-loaded in the software.This pathogen datatset is derived from the pathogen genes, and a proprietary method, so that the ectodermal response can be compared to the pathogen genes.Once the comparison is complete, the result is displayed.EPD scanning can be performed on humans and animals with consistent results.EPD scans have been performed on adults (all ages) and babies, toddlers and children.EPD scans have been performed on pregnant animals with no adverse results or outcomes to the fetus and newborn.Full Protocol and background:Pathogen Diagnosis The current SARS-COV-2 world pandemic highlights the need for widespread, fast and accurate, point of care testing, and diagnosis. Health experts across the globe agree, that the information gained from testing, is crucial in the management of the epidemiological curve, curbing the spread of the infection, reducing mortality, and optimal use and implementation of medical resources and staff.It is imperative to identify individuals with current infections, testing both symptomatic and asymptomatic individuals, to stop the human to human transmission, of the virus. To date, no country in the world has had the luxury of implementing blanket testing. Mainly because of the steep inclination of the epidemiological curve, whereby governmental and healthcare systems, cannot keep up with the rapid spread of the virus resulting in the exponential increase in symptomatic individuals requiring hospitalization and intensive care. Further challenges include logistical issues, in the distribution of testing kits to existing laboratories, setting up new facilities for sampling and testing, as demand increases. Lastly, third world countries have the added constraints of under-resourced healthcare systems and financial limitations to purchase enough molecular testing equipment.Even if blanket testing were possible, diagnosing the SARS-COV-2 infected individual accurately remains a challenge.This challenge exists for the following reasons: properties unique to the SARS-COV-2 virus (incubation period, a large portion of asymptomatic infected individuals spreading virus), the sensitivity of the RT-PCR ASSAY laboratory tests (depend on which combination of target genes are included in the various test kits, viral load present at the time of sampling, and availability of kits).A positive RT-PCR ASSAY test confirms the presence of SARS-CoV-2 viral nucleic acids (RNA), in a nasopharyngeal swab, oropharyngeal swab, or any bodily fluid or tissue. Generally the sensitivity (accuracy) of RT-PCR ASSAY for SARS-COV-2 is in the region of 80%, with high specificity. Therefore individuals with symptoms of SARS-COV-2 who test negative (up to 20%), should not be managed on the test result alone, but also taking into account epidemiological risk factors (travel history, contacts), clinical symptoms, and by retesting the patient using a different sampling site.Demonstrating seroconversion, the presence in serum of either IgM or both IgM and IgG or IgG, for SARS-COV-2, indicates a past (albeit recent past) exposure, viremia, and production of antibodies. Whether these antibodies are neutralizing and confer immunity is still being established. This type of test cannot be used to identify active infection.Due to costs and the massive logistical demands on RT-PCR ASSAY producing companies (collection kits, purification units, extraction units, reagent solution production), RT-PCR ASSAY is only used in a certain portion of the population at this stage.NICD guidelines for RT-PCR ASSAY testing published on their website https://www.nicd.ac.za/wp-content/uploads/2020/04/COVID-19-Quick-reference-v12-09.04.2020_final-1.pdf:Criteria for the person under investigation (PUI), i.e. a person to be tested for COVID-19 Persons with acute respiratory illness with sudden onset of at least one of the following: cough, sore throat, shortness of breath or fever [≥ 38°C (measured) or history of fever (subjective)] irrespective of admission status.There is an international upsurge and call for emergency research protocols being submitted and approved for clinical trials of new drugs, vaccine development studies and other technology to combat the spread and loss of life caused by this pandemic. EPD is Novel technology for pathogen diagnosisEPD scanning technology, is a completely novel technique, designed to assess the human body for the presence or absence of a pathogen.The technology implements a unique scanning method, by delivering an extremely low electrical current to the skin, and then recording the resulting physiological response to this stimulation, from the skin.Although the technique is unique, there are similarities to existing technologies that can be used to explain the modus operandi, of this novel technology. The microcurrent delivery component of the diagnostic technology can be loosely described as containing elements analogous to Microcurrent Electrical Therapy (MET) and TENS (Trans dermal electrical nervous stimulation), whereas the response component of the technology can be seen as similar to Electrocardiography (ECG), Elelctro-enchephalogram (EEG) the Electrodermal Response (EDR), and Body Impedance Assessment (BIA).All of these modalities are widely used, considered safe and have been extensively researched.The skin offers convenient access to study the nervous system, not only is it richly innervated, but it also originates from the same embryonic layer as the nervous system. It is well known that the immune system and immune response is regulated by the neurological system.EPD technology uses this physiological platform offered by the skin to access real-time information of the immune response and compares this physiological information to electronically generated pathogen molecular patterns via a proprietary algorythm.Background:Immune systemThe immune system consists of cells and proteins found in the blood, lymphatic system and tissue. These cells originate from the bone marrow, and mature in the thymus and spleen.The purpose of the vertebrate immune system is protection from invading pathogens. This protection relies on the innate immune system's ability to co-ordinate effective cell surveillance and recognition of pathogens. The ultimate goal is the eradication of the pathogen, and the production of a neutralizing antibody via the adaptive immune response, this will eventually lead to protection against disease from the same pathogen, if encountered a second time. (Zinkernagel 2002), (Chaplin 2006).Embryology of the immunes systemThe three components of the immune system originate independently of each other from different parts of the mesoderm. macrophage mesoderm, lymphatic, and hematopoietic systems splanchnic mesoderm. This co-ordination depends on chemical signalling between cells, and then recognition of pathogens by cells.Ralph Steinman, Bruce Beutler and Jules Hoffman, were awarded the 2011 Nobel Prize for Physiology in Medicine, for their contributions towards better understanding the interactions between the innate and adaptive parts of the immune response. In particular Ralph Steinman by uncovering that dendritic cells are the bridging communication link, between the cells of the innate and adaptive immune systems.Jules Hoffman and Bruce Beutler for their work on demonstrating how the innate immune system is activated. The innate immune system recognizes conserved Pathogen Associated Molecular Patterns (PAMP's) through Toll-like receptors (TLR's) expressed on the cell surfaces of immune cells. The concept of PAMP's was first coined by the immunologist CA Janeway in 1989, at a Cold Spring Harbor Symposium on Quantitative Biology, (Beutler 2011),( (Paul 2011). Steinman 2012),(Hoffman 2011) (Lemaitre 1996),(Murphy 2017). Cell communicationCells communicate in various ways. Apart from the chemical and molecular, receptor-ligand model of cellular communication, that forms the backbone of our current understanding of biochemical and immunological processes in modern science, there is also a large and rapidly growing body of evidence, that there are additional and equally effective forms of cellular communication, in biological systems.Alexander Gurwitsch was the first scientist to show that onion plant roots communicate with “mitogenic radiation”, radiation in the ultraviolet light range of the electromagnetic frequency spectrum.(Gurwisch, 1923),(Gurwisch, 1926).Professor Fritz Alexander Popp continued this line of investigation and contributed a large body of work on biophotons (photons of light in the ultraviolet and low visible light range, produced by biological systems).(Popp et al., 1994.),(Popp 1988.),(Popp and Klimek 2007).Following on Popp’s work, biophotons and cell communication have been extensively researched by other authors, and in their online book Fields of the Cell (Fels et al., 2015), their work is discussed in great detail.Irene Cosic describes in her book "The resonant recognition model of macromolecular bioactivity: theory and applications" – an alternative resonant model for cell communication, a model for digital biological activity including, it's potential applications, (Cosic 1997) this model, additional to the current model, of ligands and receptors, describing biological processes.Robert Becker the American orthopaedic surgeon, answered the question of how a lizard perfectly grows back an amputated limb. By implementing meticulous research procedures, he showed that following trauma, tissue electrical fields produced at the site of injury, provides an information and communication platform, for cells, to regenerate tissue and amputated limbs to their original specifications. (Becker and Snelden 1998).In his paper “The Electrical Embryo”, Colin Lowry discusses that electrical fields are essential in embryonic growth, pattern and shape (Lowry 1999).Cells react to electrical stimulation, fields and frequenciesIt has also been shown that electrical stimulation, affects cell membrane receptors and ATP production. (Seegers at al. 2001) demonstrated that a microcurrent DC electrical field of less than 150Hz, interacts with HeLa cell receptors, producing a 50-fold increase in ATP in vitro, and a 163% increase in vivo in plasma.Similar findings of electric frequencies activating receptors were published by (Wolf-Goldberg et al. 2013.) showing that Epidermal Growth Factor Receptor (EGFR) activation can be induced by applying a short train of pulsed low strength electric field (LEF) frequencies with 10v/cm pulse-width for 180µs, at 500Hz, for 2min.The idea of using electronic devices, including biosensors to detect pathogenic bacteria is not new and this concept was discussed comprehensively in a review article by (Ivnitski et al.1999). Technological advances since then, including nanotechnology, has opened new doors and new diagnostic possibilities.(Patolski et al. 2004.) using nanowire field resistance, demonstrated that sensitive detection of a single virus is possible by coating a nanowire with Influenza specific antibodies and then adding an Influenza virus (antigen) to this coated nanowire, conductance was observed and a flow of current was demonstrated upon molecular docking, with the specific antibody binding to the virus.Similarly using a slightly different technique, namely impedance spectroscopy, (Abdelghani 2011) demonstrated sensitive detection of the Rabies Virus.The Nervous SystemConsists of the Central end Peripheral nervous system. The central nervous system – the brain and spine, and the peripheral nervous system consists of the somatic and autonomic nervous system. The autonomic nervous system (ANS) is divided in the Sympathetic (SNS) and Parasympathetic nervous system (PNS). Embryologically the nervous system develops from the ectoderm. (Sadler 2011).Immunity is regulated by the Neurological systemThe nervous and immune systems are in constant two-way communication.Our current knowledge and understanding of the immune response, and the regulator thereof, through neuro immune interaction, is based on a large body of scientific work done over the last decade.The primary function of the autonomic nervous system (ANS) is to help the body adapt to internal and external environmental stimuli and demands, by maintaining a healthy balance. (Beissner et al. 2013).The sympathetic nervous system stimulates increased metabolic output.This is known as sympathetic arousal and is characterized by elevated heart rate, blood pressure, sweating and, shunting of blood from the internal gastrointestinal reservoir to the heart, brain and, skeletal muscle, in preparation of a "fight or flight" response.The parasympathetic nervous system effects the opposite, by inducing conservation of metabolic energy and restoration.The brain and immune system are in constant two-way communication to maintain health, and the sympathetic nervous system plays an integrative role in the regulation of the immune response. This physiological equilibrium between the two systems is regulated by two pathways.Centrally, via the Hypothalamic Pituitary Axis Adrenal Corticotropin Pathway (HPA-CRH) and peripherally via the autonomic nervous system (ANS), which in turn is comprised of the sympathetic (SNS) and parasympathetic (PNS) nervous systems. (Dantzer 2019).Loeper and Crouzon demonstrated in 1904 that subcutaneous injection of epinephrine (adrenaline) causes a marked leucocytosis in humans.Primary and secondary lymphoid organs receive substantial sympathetic innervation.With sympathetic nervous system activation, neurotransmitters are released from sympathetic nerve endings in the lymphoid organs, they then bind to specific receptors on lymphatic cells, not only affecting cell traffic, division and, circulation, but also the production of cytokines. This may modulate the type of immune response to either a Th1 cell or Th2 cell type. (Pavlov et al. 2003), (Kin and Sanders 2006).Studying the sympathetic nervous system and particularly the sympathetic response has been ongoing since the 1970's using a variety of biofeedback systems.The most widely studied and used of these methods must be the Galvanic Skin Response, otherwise known as Skin Conductance or Electro Dermal Activity (EDA), (Dawson 2007).Furthermore, T-lymphocytes have also been found to have B2 adrenergic receptors), indicating a neurological regulation of the immune response. (Fan and Wang 2009).The Integumetary systemConsists of the skin and its appendages – hair and nails. The purpose of this system is to protect the body from various kinds of damage, such as loss of water or external harm. Apart from these it also has other functions; thermoregulation, waterproofing, protecting deeper tissues, and waste excretion. The skin is richly innervated with sensory receptors for pain, temperature, pressure and sensation. Embryologically the skin originates from the ectoderm. (Langman’s Essential Medical Embryology, 2011).The skin is the largest organ in the human body, the first defence barrier between the human body and the external environment offering protection against pathogens, chemicals, toxins, and is also richly innervated with about 2500 per cm² nerve endings in the fingertips. (McBride and Schmorrow 2005).Lastly, the integumentary system synthesizes vitamin D.The skin utilizes solar energy which is transmitted as electromagnetic waves. These waves are classified according to their wavelength and frequency. The skin makes use of these properties of the electromagnetic spectrum to produce Vitamin D. (Cavalcanti Soriano Coutinho et al. 2019)Microcurrent electrical therapy (MET)Microcurrent is used for wound healing and pain relief. (Wirsing et al.2013)Application of subsensory micro-amperage current (300 to 500 micro amps pulsed at 0.1 to 680 Hz) to tissue, increases cellular output of ATP. (Seegers et al. 2002)These currents compliment and augment the physiological tissue electrical fields produced at the site of injury which provides an information and communication platform, for cells, to regenerate tissue and amputated limbs to their original specifications. (Becker and Snelden 1998).TensThis technology is widely used for pain management.Application of milliamp current, to blocks pain messages, relayed by sensory nerve fibres, A beta (Aβ), (afferent: to the brain), thereby reducing the ‘c’ fibre (efferent: from the brain) noxious stimulus transmission (pain sensation). The two primary pain relief mechanisms which are targeted are: the “Pain Gate Mechanism” and the “Endogenous Opioid System”. Endorphins (endogenous opoids) are produced in the brain, upon peripheral nervous stimulation.The current intensity (A) (strength) will typically be in the range of 0 - 80 mA up to 100mA. The current is pulsed (the pulse rate or frequency (B) will normally be variable from about2 – 150 Hz. (Vance et al. 2014.)Electrocardiogram (ECG)The heart is a self-powered pump, cardiac pacemaker cells, produce a pace-making current (action potentials) of up to about -50 millivolts, with an amplitude of about 5 nano amperes. This current travels from pacemaker cells via conducting fibers, through the heart muscles, triggering muscle contractions (pumping actions). This current can be measured from the skin with electrodes, (the electrolyte rich fluid of the body, is a good conductor of current). This current of about 1 millivolt, is displayed as a graph the ECG – and represents the sum of all the electrical potentials generated by pacemaker cells at any given time. (Fye 1994)Electroenchephalogram (EEG)Neurons communicate electrically and chemically. Mammalian neurons, produce action potentials of up to about -55 millivolts, with an amplitude of about 1 nano ampere. This can be seen as units of power or watts, or 1.05 x 10^-10 W / neuron. There are billions of neurons in a human brain. Total brain wattage can be calculated as, (1.05 x 10^-10 W / neuron) x (800 million neurons / brain) = 0.085 Watts / brain. The electrical activity in the brain can be recorded via scalp electrodes and displayed on a graph like an ECG, where voltage is plotted against time. The general voltage range of the scalp EEG lie between 10 and 100 µV.(Nunez 2006). (Bronzino, 1999).Electro Dermal Activity (EDA)Electrodermal activity (EDA) is the umbrella term used for defining autonomic changes in the electrical properties of the skin. Historically the two principles used in modern-day skin conductance were discovered by Féré in 1888, who demonstrated decreases in skin resistance with different sensory stimuli, and the Russian physiologist Tarchanoff in 1890, who showed changes in electrical conduction between two skin electrodes during sensory stimulation.(Fere 1888),(Dawson 2007).Skin Conductance or EDA has been shown to provide a sensitive index of the status of the sympathetic nervous system. (Critchley et al., 2000.), (Critchley, 2002).Sweat glands or eccrine glands (sudomotor) are innervated by sympathetic post-ganglionic fibers, consisting of non- myelinated class C fibers. Activation of these nerve fibres modulates sweat secretion. (Figure 1). EDA "taps" into these nerve fibres, and therefore into the sympathetic nervous systemCorrelation between EDA and functional MRI has shown an extensive network of the SNS. (Beissner et al., 2013.), (Critchley et al., 2002). A recent study at Harvard University's Massachusetts Institute of Technology (MIT) by Professor Rozalind Pickard, whilst investigating emotional responses in Autistic Children, revealed an unexpected finding, that EDA can predict the onset of Sudden Death Epilepsy (SUDEP) 24 hours prior the event. SUDEP is a life-threatening condition with cardiac arrest, following post-ictal generalized electrical suppression (Sarkis et al., 2013).Following this discovery, the EMBRACE bracelet warning system was developed. (Poh, 2011.),(www.empatica.com).Bioelectrical Impedance Analysis (BIA) and EPD scan measuring techniqueBioelectrical Impedance Analysis (BIA) takes advantage of the conductive properties of the body, and is a method extensively used in studies assessing body composition, water, fat, muscle. (Kyle 2004),(Mialich et al. 2014).The body’s response to electric current, is similar to an RC circuit, where body fluids (intracellular water (ICW) and extracellular water (ECW)) are represented by resistors and cell walls by capacitors. The impedance measurement is generally measured from the wrist to the contralateral ankle and uses either two or four electrodes. A small current on the order of 1-10 μA is passed between two electrodes, and the voltage is measured between the same (for a two electrode configuration) or between the other two electrodesFat cells have very high impedance or resistance compared to muscle cells, organ cells, extracellular components (including connective tissue). Therefore, electric current will mostly propagate through the fat-free body mass and not the fatty tissue. This means that BIA measures the volume of fat-free mass.EPD technology a combines existing and novel technology for pathogen diagnosis.EPD technology is utilizes the principles of electric stimulation and the resulting response thereto, to investigate the real-time immune response to pathogens.However EPD technology goes further in that it can identify the presence or absence of a pathogen in the mammalian body from these readings.Frequencies are applied in the range of 10 kHz to 500 kHz in a proprietary manner. The body’s reaction is measured and statistically analyzed using propriatory algorythms.This measuring technique uses voltages in the range of 6-volt peak to peak (similar to a line-up of 4 penlight batteries) generating a body current of approximately 500 micro-amp (500 millionth of one amp), at applied frequencies. This well below all safety standards, and is proven to be safe for human subjects. (ICNIRP,1998).Molecular identification of PathogensWatson and Crick received the Nobel prize in 1962 for "for their discoveries concerning the molecular structure of nucleic acids and its significance for information transfer in living material." Kerry Mullis in recognition of his invention of the polymerase chain reaction technique, shared the Nobel Prize in 1993 in Chemistry with Michael Smith. (Crick 1962),(Mullis 1993),(Watson 1962).These two scientific milestones paved the way for modern day molecular biology and medicine and our understanding of diseases.It is now possible to sequence the genomes of pathogens and their proteins rapidly, and this information is uploaded to large open domain databases.EPD technology makes use of this information.For the purpose of the Xprise competition, the complete sequenced genome, all genes of SARS-CoV-2 virus will be implemented. However due to the unique properties of EPD technology, it is possible to target not only the viral genome, but also the proteome, and immune response including, lymphocyte antigens, cytokine production, to the viral infection. Cytokines identification, and individual proteins testing are scarce resources and not widely available for data validation in South Africa. EPD technology makes use of an AI model taking all of the above into account.This approach has been used in a Pilot Study using EPD technology on Bovines infected with Mycobacterium Bovis.(Michel et al.2018)SafetyThe safety of bioelectrical instrumentation is assessed by two parameters. One is the aspect of electrical isolation from ground potentials for the subject. The second is the definition of what is a harmless current vs. frequency that can be deliberately introduced into the subject. There are few references that have explicitly established the standards for what is a safe subject current and frequency. (Geddes andBaker, 1975) in Applied Biomedical Instrumentation, describe the threshold of electrical perception of alternating currents of varying frequency.There have been many applications of electrical impedanceat frequencies from 10 KHz to 5 MHz that have been introduced to critical human organs. (Nyboer and Kornmesser, 1970) applied an impedance plethysmograph (designed and built by Rudolph J. Liedtke) to the area of the uterus, to monitor pregnancy labor movements. There were no reported abnormalities after these observations using this method. The instruments that were used had a frequency of 100 KHz (crystal controlled) at approximately 3 milliamps. (Bishop and Nyboer,1970) applied the same instrument directly to the eye with an electrode array, configured in a contact lens, with no ill effects at this frequency and current (100 KHz at 3 milliamps).The EPD frequency in the range of 10 kHz to 600 kHz. This measured body reaction, is mathematically analyzed to extract information, that seems to bear a relationship between certain frequencies, and viruses, that have activated the body’s immune system at the time of measurement.This measuring technique, uses voltages in the range of 6 volt peak to peak (similar to a line-up of 4 penlight batteries) generating a body current of approximately 500 micro-amp (500 millionth of one amp), at applied frequencies.This well below all safety standards, and is proven to be safe for human subjects.(ICNIRP,1998).Body exposure to applied electrical signals: EPD Tracer·Electrical Current used for measurement : 100 to 800 micro-amp induced by applying up to 6 volts·Applied Frequency: 10 to 600 Kilo Hertz·Emf radiation : None – direct application of measuring currentEDR Electro dermal activityElectrical Current used for measurement : NoneSensor Frequency: 4 HertzEmf radiation : None (BIA) Body Impedance Assessment·Electrical Current used for measurement : 100 to 800 micro-amp using up to 6 volts·Applied Frequency: 2100 to 600 Kilo Hertz·Emf radiation : None – direct application of measuring currentTENS Trans dermal electrical nervous stimulation·Electrical Current used for measurement : up to 50 milli-amp using up to 9 volts·Applied Frequency: 250 Hertz pulses with pulse duration .01 to1 milli -seconds·Emf radiation : None – direct application of measuring current.EPD Modus Opperandi:Introduction Gene TargetsAll sequenced genes of the SARS-CoV-2 virus.Note: Make sure the SARS-CoV-2 virus text file is correctly uploaded into the EPD software.Equipment-EPD scanner-Transmitting electrode-Receiving electrode-Laptop with EPD software-Patient-Table-Chair x 2-PPE-Alcohol based surface disinfectant-Hand sanitizerEPD software-EPD 101ProcedureStep 1:Switch on EPD scan unit and PC, connect EPD scan unit to PC.Note: Make sure both PC and EPD scanner are on, and connected.Step 2:Open EPD software.Note: Make sure both PC and EPD scanner are on, and connected.Step 3:Enter patient details into EPD software.Note: Step 4:Connect receiving electrode to LT middle finger terminal phalange of patient. Give patient transmitting electrode to hold in RT hand.Note: Clean electrodes with alcohol based disinfectant. Make sure hands are clean and skin is dry.Step 5:Commence scan.Note: Make sure the EPD software reads the EPD scan machine.Step 6:Read the result.Note:Step 7:Communicate the result.Note: TroubleshootingIf problems getting scan to initiate:Check that scanner is on and connected. Make sure skin is clean and dry.Make sure probes are clean and dry.Time TakenFrom the time that the scan is commenced, to final result takes around 8 minutes. Anticipated ResultsAccurate diagnosis of the presence or absence of SARS-CoV-2 virus.References:ABDELGHANI, A.,2011.Electrochemical Biosensors for Virus Detection, Biosensors for Health, Environment and Biosecurity, ed. Prof.Serra, P.A.,Retrieved from:InTechhttp://www.intechopen.com/books/biosensors-for-health-environment-and-biosecurity/electrochemical-biosensors-for-virus-detection.BECKER, R.O.,and SELDEN, G.,1998The Body Electric: Electromagnetism and the Foundation of Life Harper Paperbacks New YorkBEISSNER, F., MEISSNER, K., BAR, K.J., NAPADOW, N., 2013.The Autonomic Brain An Activation Likelihood Estimation Meta-Analysis For Central Processing of Autonomic FunctionThe Journal of Neuroscience, 33, (25),10503 -10511.BEUTLER.B.,2011,Nobel Lecture. NobelPrize.org. Nobel Media AB 2020. Sat. 5 Sep 2020. https://www.nobelprize.org/prizes/medicine/2011/beutler/lecture/BEUTLER, B., GREENWALD, D., HULMES, J.D., CHANG, M.,MATHISON, J.,ULEVITCH, R.,CERAMI, A.,1985.Identity of tumour necrosis factor and the macrophage-secreted factor cachectin.Nature 316,(6028),552-558.BEUTLER, B., HOEBE, K., DU, X., ULEVITCH, R.J.,2003.How we detect microbes and respond to them: the Toll-like receptors and their transducers. Journal of Leukocyte Biology,74,479-85.BISHOP, S., NYBOER,J.,1970.Electrical impedance of the anterior eye chamber.Annals of the New York Academy of Science,170, (2),793-800.BRONZINO, J.D.Principles of ElectroencephalographyThe Biomedical Engineering Handbook19992nd Edition, ed, BRONZINO,D.CRC Press CAVALCANTI SORIANO COUTINHO.R,  FEITOSA DOS SANTOS.A , GOMES DA COSTA.J, DIAS VANDERLEI.A, 2019Sun exposure, skin lesions and vitamin D production: evaluation in a population of fishermenAn Bras Dermatol. Jul 29;94(3):279-286CHAPLIN, D.D, 2006.Overview of the human immune responseThe Journal of allergy and clinical immunology, 117,(2 Suppl Mini-Primer),S430-5.CRICK.F, 1962, Nobel Lecture. NobelPrize.org. Nobel Media AB 2020. Sun. 6 Sep 2020. https://www.nobelprize.org/prizes/medicine/1962/crick/lecture/CRITCHLEY, H.D.,2002.Electrodermal Responses: What Happens In The BrainThe Neuroscientist,8,(2),132-142.CRITCHLEY, H.D., ELLIOT, R., MATHIAS, C.J., DOLAN, R.J., 2000. Neural Activity Relating to Generation and Representation of Galvanic Skin Conductance Responses:A Functional Magnetic Resonance Imageing Study.The Journal of Neuroscience 20,(8),3033-3040.COSIC, I.,1997The resonant recognition model of macromolecular bioactivity : theory and applicationsBasel: BirkhäuserDAWSON, M.E., The Electrodermal System2007Handbook of Psychophysiology 3rd EditionCacioppo,J.T, Tassinary,L.G., Berntson,G.,ed,Cambridge University Press New YorkDANTZER.R, 2017Neuroimmune Interactions:from the brain to the immune system and vice versaPhysiol Rev 98: 477–504FAN, X., WAN, Y., 2009.β2 Adrenergic receptor on T Lymphocytes and its Clinical implications Progress in Natural Science,19,17-23.FELS,D, CIFRA,M, SKOLKMANN.F.2015Fields of the CellResearch SignpostFÉRÉ,C.,1888.Note on changes in electrical resistance under the effect of sensory stimulation and emotionComptes Rendus de Seances de la Societe de la Biologie,9,(5),217-219.FYE.W.B, 1994 A History of the origin, evolution, and impact of electrocardiographyThe American Journal of Cardiology, Volume 73, ISSUE 13, P937-949, May 1515GURWITSCH, A.G.1926. Das Problem der Zellteilung, physiologisch betrachtetProtoplasma 1/1, 473-475.GURWITSCH, A.G., 1923.Die Natur des spezifischen Erregers der ZellteilungArchiv für Entwicklungsmechanik der Organismen,100,(1-2), 11–40.HOFFER, E.C., MEADOR, C.K., SIMPSON, D.C, 1969.Applied Physiology, 27, 531. HOFFMANN J.A,2011, Nobel Lecture. NobelPrize.org. Nobel Media AB 2020. Mon. 7 Sep 2020. https://www.nobelprize.org/prizes/medicine/2011/hoffmann/lecture/INTERNATIONAL COMMISION ON NON- IONIZING RADIATION PROTECTION, (ICNIRP) GUIDELINES.,1998.Health Physics 74 (4):494‐522; IVNITSKI,D., ABDEL-HAMID,I.,ATANASOV,P.,WILKINS,E.,1999Biosensors for Detection of Pathogenic BacteriaBiosensors & Bioelectronics,14,599-624.KIN, N. W., SANDERS, V.M., 2006.It takes nerve to tell T and B cells what to do.Journal of Leukocyte Biology, 79, 1093 – 1104.KUCERA, P., GOLDENBERG, Z., KRUCA, E., 2004.Sympathetic Skin response: review of the method and its clinical useBratisl Lek Listy, 105,(3),105-116.KYLE, U.G., BOSAEUS, I., DE LORENZO, A.D., DEURENBERG,P., ELIA, M., GOMEZ, J. M., HEITMANN, B. L., KENT – SMITH, L., MELCHIOR, J-C., PIRLICH, M., SCHARFETTERK, H., SCHOLS, A. M.W.J., PICHARD, C, Composition of the ESPEN Working GroupESPEN GUIDELINESBioelectrical impedance analysis: Part I: review of principles and methods Clinical Nutrition (2004) 23, 1226–1243LEMAITRE, B., NICHOLAS, M., MICHAUT, L., REICHART, J-M., HOFFMAN, J.A., 1996. The Dorsoventral Regulatory Gene Cassette spätzle/Toll/cactus Controls the Potent Antifungal Response in Drosophila AdultsCell,86, 973-983.LIEDTKE, R.J., 1997Principles of Bioelectrical Impedance AnalysisRetrieved from: http://rjlsystems.comLOEPER, M., CROUZON, O., 1904.L’action de l’adrenaline sur le sang. Arch Med Exp Anat Pathol, 16,83–108.LOWRY, C.The Electric Embryo: How Electric Fields Mold the Embryo's Growth Pattern and Shape199921st CENTURY SCIENCE & TECHNOLOGYSpringerMCBRIDE, D.K, SCHMORROW, D.,Quantifying Human Information Processing)2005: Lexington Books MIALICH, M. S., FACCIOLI SICCHIERI, J. M., JORDAO JNR, A. A., 2014.Analysis of Body Composition: A Critical Review of the Use of Bioelectrical Impedance AnalysisInternational Journal of Clinical Nutrition,2(1),1-10.Mullis.K, 1993, Nobel Lecture. NobelPrize.org. Nobel Media AB 2020. Sat. 5 Sep 2020. https://www.nobelprize.org/prizes/chemistry/1993/mullis/lecture/MURPHY.K, WEAVER, C, JANEWAY.C, 2017Janeway's Immunobiology9th editionGarland Science New York NYBOER, J., KORNMESSER, G.D., 1970.Electrical Impedance of the abdomen during Maternal LaborAnnals of the New York Academy of Sciences International Conference on Bioelectric Impedance, 170, Pages 801–803.PAUL.W.E.,2011.Bridging Innate and Adaptive ImmunityCell, 147,(6),1212-1215.PATOLSKY, F., ZHENG, G., HAYDEN, O., LAKADAMYALL, M., ZHUANG, X., LIEBER, C.M., 2004.Electrical detection of single virusesPNAS ,101,(39),14017-14020.PAVLOV, V.A., WANG, H., CZURA, C.J, FRIEDMAN, S.G., TRACEY, K.J., 2003.The Cholinergic Anti Inflammatory Pathway: A Missing Link in Neuro-immunomodulationMolecular Medicine, 2003,(9), 5-8.POH, M-Z.,2011.Continuous Assessment of Epileptic Seizures with Wrist-worn Biosensors(Doctoral Thesis)Retrieved from: https://dspace.mit.edu/handle/1721.1/68456POPP, F. A., 1988. Biophoton emissionExperientia ,Basel, 44, 543–544. POPP, F. A., CHANG, J.-J., GU, Q., Ho, M.-W.,Nonsubstantial biocommunication in terms of Dicke's theory BIOELECTRODYNAMICS AND BIOCOMMUNICATIONHo, M.-W., Popp, F. A., Warnke, U., ed, 1994:World Scientific, Singapore. POPP, F. A. & KLIMEK, W., Photon Sucking as an Essential Principle of Biological Regulation BIOPHOTONICS AND CHOERENT SYSTEMS IN BIOLOGY Beloussov, L. V., Voeikov, V. L., Martynyuk, V. S, ed,2007: Springer, New York POPP, F. A. & Yan, Y., 2002. Delayed luminescence of biological systems in terms of coherent states. Physics Letters, A, 293, 93–97. SADLER,T.W.LANGMAN’S ESSENTIAL MEDICAL EMBRIOLOGY201112th Edition Lippincot Williams and Wilkins SARKIS, R., THOME-SOUZA, S., POH, M., KLEHM, J., VANNAN, D., MADSEN, J., PICKARD, R., LODDENKEMPER,T., REINSBERGER,T., 2013.Autonomic changes following generalised tonic clonic seizure: and analysis of adult and paediatric patients with epilepsy. American Epilepsy Society Annual General Meeting, Washington DC, Abstract 2.069SCHOLKMANN,F., FELS,D., CHIFRA,D., 2013.Non-chemical and non-contact cell to cell communication: a short reviewAmerican Journal of Translational Research 5,(6),586-593.SEEGERS, J.C., LOTTERING, M.L., JOUBERT, A.M., JOUBERT,F., KOORTS, A., ENGELBRECHT, C.A., VAN PAPENDORP, D.H., 2002.A pulsed DC electric field affects P2-purinergic receptor functions by altering the ATP levels in in vitro and in vivo systemsMedical Hypotheses, 58,(2),171-176.STEINMAN,R., 2011Nobel Lecture. NobelPrize.org. Nobel Media AB 2020. Sun. 6 Sep 2020. https://www.nobelprize.org/prizes/medicine/2011/steinman/lectureSTEINMANN.R. COHN, Z.A.,1973.Identification of a Novel Cell Type in Peripheral Lymphoid Organs of MiceThe Journal of Experimental Medicine, 1367,1142-1162.TARCHANOFF,J., 1890.Galvanic phenomena in the human skin during stimulation of the sensory organs and during various forms of mental activityPflügers Archiv für die gesammte Physiologie für des Menschen und der Tiere, 46,46-55. WATSON.J, 1962, Nobel Lecture. NobelPrize.org. Nobel Media AB 2020. Sat. 5 Sep 2020. https://www.nobelprize.org/prizes/medicine/1962/watson/lecture/WIRSING.P.G., HABROM.A.D., ZEHNDER.T.M, FRIEDLI.S., Blatti.M,2013Wireless micro current stimulation – an innovative electrical stimulation method for the treatment of patients with leg and diabetic foot ulcersInternational Wound Journal · December 2013WOLF-GOLDBERG, T., BARBUL, A., BEN-DOV, N., KORENSTEIN, R., 2013.Low electric fields induce ligand-independant activation of EGF receptor and activation of ERK via electrochemical activation of H+ and ROSBiochimica et Biophuiica Acta 1833, 1396-1408.VANCE.C.G.T, DAILEY.D.L, RAKEL.B.A, SLUKA.K.A 2014Using TENS for pain control: the state of the evidencePain Manag. May;4(3):197-209.ZINKERNAGEL,R.M., 2002mmunological Reviews 185,103–125.

Proper citation: Ilse Pretorius 2020. EPD Electronic Pathogen Detection. protocols.io dx.doi.org/10.17504/protocols.io.bkgzktx6 Copy   

•   Source: Protocols.io

Authors: Devin Willis
Group: XPRIZE Rapid Covid Testing, RNAPath

Proper citation: Devin Willis 2020. RNAPath Target Identification via Fluorescent Hybridization. protocols.io dx.doi.org/10.17504/protocols.io.bk37kyrn Copy   

•   Source: Protocols.io

Authors: Rob Parks, Barton Haynes
Group: XPRIZE Rapid Covid Testing

Proper citation: Rob Parks, Barton Haynes 2020. SARS-CoV-2 Antigen Detection ELISA. protocols.io dx.doi.org/10.17504/protocols.io.bk2ckyaw Copy   

•   Source: Protocols.io

Authors: yliu
Group: OnsiteGene 1, XPRIZE Rapid Covid Testing
Summary: This OnsiteGene protocol is designed for testing the normal saliva samples with nucleic acid extraction. A normal saliva collection tube is used for the collection. The protocol uses the Star Array® Hi-SenseTM COVID-19 Molecular Testing Kit 1.0 in the one-step real-time RT-qPCR test assay to qualitatively detect RNA from SARS-CoV-2 virus in the human respiratory specimen. It combines the reverse transcription technology and real-time PCR method to provide accurate detection of the SARS-CoV-2 coronavirus. The Star Array® magnetic beads RNA extraction instrument and kit can automatically extract and purify the RNA for 32 samples in 6 minutes. It is ideal for the saliva samples that contain high amount of PCR inhibitors and can't be processed by the direct amplification. The triplex fluorescence design of the kit simultaneously detects the N1 and Orf1ab genes of the virus and the human RNase P gene as an internal control to ensure the sample quality. This protocol uses the Star Array® XDiveTM Superfast Real-Time PCR instrument to perform 40-cycle PCR in 8 minutes, and can test up to 16 samples or controls in each run. The total protocol time from sample collection to data interpretation is less than 15 minutes.The sample collection steps is adapted from the published protocol:https://www.protocols.io/view/saliva-collection-and-rna-extraction-for-sars-cov-bh6mj9c6

Proper citation: yliu 2020. OnsiteGene 4 Protocol Saliva Extract. protocols.io dx.doi.org/10.17504/protocols.io.bpjemkje Copy   

•   Source: Protocols.io

Authors: Jon Arizti-Sanz, Catherine A. Freije, Chloe K. Boehm, Sameed M. Siddiqui, Allen M. Goodman, Tinna-Solveig F. Kosoko-Thoroddsen, A'Doriann Y. Bradley, Jeremy Johnson, Pardis C. Sabeti, Cameron Myhrvold
Group: XPRIZE Rapid Covid Testing
Summary: This protocol describes how to perform a SHINE paper-based assay to detect SARS-CoV-2 RNA from a self-collected saliva sample. This protocol is intended for in-home use. All enzymatic components are provided as a single-test freeze-dried pellet for shelf-stable storage, and all steps of the protocol are performed at ambient temperature. The protocol presented here is an improved version of the method presented in Arizti-Sanz J*, Freije CA*, et al. Integrated sample inactivation, amplification, and Cas13-based detection of SARS-CoV-2. bioRxiv (2020).

Proper citation: Jon Arizti-Sanz, Catherine A. Freije, Chloe K. Boehm, Sameed M. Siddiqui, Allen M. Goodman, Tinna-Solveig F. Kosoko-Thoroddsen, A'Doriann Y. Bradley, Jeremy Johnson, Pardis C. Sabeti, Cameron Myhrvold 2020. XPRIZE SHINE - Paper-based SARS-CoV-2 Saliva Test. protocols.io dx.doi.org/10.17504/protocols.io.bk2fkybn Copy   

•   Source: Protocols.io

Authors: philippe.bechtold
Group: XPRIZE Rapid Covid Testing

Proper citation: philippe.bechtold 2020. Sample preparation. protocols.io dx.doi.org/10.17504/protocols.io.bjygkptw Copy   

•   Source: Protocols.io

Authors: Chantal Vogels, Doug E. Brackney, Chaney Kalinich, Isabel Ott, Nathan Grubaugh, Anne Wyllie
Group: Coronavirus Method Development Community, XPRIZE Rapid Covid Testing
Summary: SalivaDirect is an RNA-extraction free, dual-plexed RT-qPCR method for SARS-CoV-2 detection. It can be broadly implemented as it (1) does not require saliva collection tubes containing preservatives, (2) does not require specialized equipment for RNA extraction, and (3) is validated for use with products from multiple vendors. Thus, the simplicity and flexibility of SalivaDirect means that it is not as affected by supply chain bottlenecks as some other assays. Our method is RNA-extraction free which enables testing of low volume and minimally processed saliva in dual-plexed RT-qPCR for SARS-CoV-2 detection. Saliva will be treated with proteinase K followed by a heat inactivation step, and is then directly used as input in the dual-plexed RT-qPCR test. Our aim was not to design new primers and probes for RT-qPCR testing, but rather to use validated primer and probe sets (N1 and RP) developed by the US CDC. The human Ribonuclease P (RP) probe was modified with a different fluorophore so that the primer/probe set could be combined in a dualplex assay, reducing the number of tests to 1 assay with 2 sets.Version 2 includes: Optimized thermocycler conditionsLocally validated alternative options for Proteinase K, RT-qPCR master mix, and thermocyclersUse of 8-strip tubes for sample processing step, due to contamination issues in 96-well plates.Version 3 has been updated to remove steps for sample self-collection.Version 4 has updated Ct thresholds for the ABI 7500 Fast Dx.

Proper citation: Chantal Vogels, Doug E. Brackney, Chaney Kalinich, Isabel Ott, Nathan Grubaugh, Anne Wyllie 2020. SalivaDirect: RNA extraction-free SARS-CoV-2 diagnostics. protocols.io dx.doi.org/10.17504/protocols.io.bjswknfe Copy   

•   Source: Protocols.io