WHotLAMP: A simple, inexpensive, and sensitive molecular test for the detection of SARS-CoV-2 in saliva

Despite the development of effective vaccines against SARS-CoV-2, epidemiological control of the virus is still challenging due to slow vaccine rollouts, incomplete vaccine protection to current and emerging variants, and unwillingness to get vaccinated. Therefore, frequent testing of individuals to identify early SARS-CoV-2 infections, contact-tracing and isolation strategies remain crucial to mitigate viral spread. Here, we describe WHotLAMP, a rapid molecular test to detect SARS-CoV-2 in saliva. WHotLAMP is simple to use, highly sensitive (3.6 viral RNA copies per microliter of saliva) and specific, as well as inexpensive, making it ideal for frequent screening. Moreover, WHotLAMP does not require harsh chemicals or specialized equipment and thus can be performed in point-of-care settings, and may also be adapted for resource-limited environments or home use. While applied here to SARS-CoV-2, WHotLAMP can be easily modified to detect other pathogens, making it adaptable for other diagnostic assays, including for use in future outbreaks.

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Mobilefuge: A low-cost, portable, open source, 3D-printed centrifuge that can be used for purification of saliva samples for SARS-CoV2 detection

10.1101/2021.01.06.21249280 ◽

2021 ◽

Author(s):

Mohini Bhupathi ◽

Ganga Chinna Rao Devarapu

Keyword(s):

Low Cost ◽

Power Supplies ◽

Testing Methods ◽

Resource Limited Settings ◽

Resource Limited ◽

Viral Particles ◽

Nasal Swabs ◽

3D Printed ◽

Frequent Testing ◽

The Cost

One of the best ways to contain the spread of COVID-19 is frequent testing of as many people as possible and timely isolation of uninfected personnel from infected personnel. However, the cost of massive testing is affordable in many countries. The existing technologies might not be scalable to offer affordable testing for millions of people. To address this issue, novel testing methods based on Loop-Mediated Isothermal Amplification (LAMP) were proposed that are more sensitive, require less reagents and can work with saliva samples instead of more tedious nasal swabs. As a result, LAMP based protocols can make it possible to drive the cost down to one dollar per test. These LAMP based methods require a centrifuge device, mostly for separation of viral particles from reaction inhibitors in saliva samples. However, centrifuge is neither accessible nor affordable in many resource limited settings, especially during this pandemic situation when normal supply chains are heavily disrupted. To overcome these challenges, we invented a low-cost centrifuge that can be useful for carrying out low-cost LAMP based detection of SARS-Cov2 virus in saliva. The 3D printed centrifuge (Mobilefuge) is portable, robust, stable, safe, easy to build and operate. The Mobilefuge doesn't require soldering or programming skills and can be built without any specialised equipment, yet practical enough for high throughput use. More importantly, Mobilefuge can be powered from widely available USB ports, including mobile phones and associated power supplies. This allows the Mobilefuge to be used even in off-grid and resource limited settings. We believe that our invention will aid the efforts to contain the spread of COVID-19 by lowering the costs of testing equipment. Apart from the COVID-19 testing, the Mobilefuge can have applications in the field of biomedical research and diagnostics.

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A Single and Two-Stage, Closed-Tube, Molecular Test for the 2019 Novel Coronavirus (COVID-19) at Home, Clinic, and Points of Entry

10.26434/chemrxiv.11860137 ◽

2020 ◽

Cited By ~ 12

Author(s):

Mohamed El-Tholotha ◽

Haim H. Bau ◽

Jinzhao Song

Keyword(s):

Point Of Care ◽

Isothermal Amplification ◽

Molecular Test ◽

Rt Pcr ◽

Two Stage ◽

Highly Sensitive ◽

Closed Tube ◽

The Usa ◽

Novel Coronavirus ◽

At Home

<p>The 2019 novel coronavirus (COVID-19) is a newly emerged strain that has never been found in humans before. At present, the laboratory-based reverse transcription-polymerase chain reaction (RT-PCR) is the main method to confirm COVID-19 infection. The intensification of the COVID-19 epidemic overwhelms limited clinical resources in particular, but not only, in developing countries, resulting in many patients not being tested for the infection and in large queues of potentially infected individuals waiting to be tested while providing a breeding ground for the disease. We describe here a rapid, highly sensitive, point-of-care, molecular test amenable for use at home, in the clinic, and at points of entry by minimally trained individuals and with minimal instrumentation. Our test is based on loop mediated isothermal amplification (COVID-19 LAMP) and for higher sensitivity on nested nucleic acid, two stage isothermal amplification (COVID-19 Penn-RAMP). Both tests can be carried out in closed tubes with either fluorescence or colorimetric (e.g., leuco crystal violet LCV) detection. COVID-19 LAMP performs on par with COVID-19 RT-PCR. COVID-19 RAMP has 10 fold better sensitivity than COVID-19 LAMP and COVID-19 RT-PCR when testing purified targets and 100 times better sensitivity than COVID-19 LAMP and COVID-19 RT-PCR when testing rapidly prepared sample mimics. Due to fortunate scarcity of COVID-19 infections in the USA, we were not able to test our assays and methods with patient samples. We hope that such tests will be carried out by colleagues in impacted countries. Our Closed-Tube Penn-RAMP has the potential to significantly reduce false negatives while being amenable to use with minimal instrumentation and training. </p>

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A Single and Two-Stage, Closed-Tube, Molecular Test for the 2019 Novel Coronavirus (COVID-19) at Home, Clinic, and Points of Entry

10.26434/chemrxiv.11860137.v1 ◽

2020 ◽

Cited By ~ 28

Author(s):

Mohamed El-Tholotha ◽

Haim H. Bau ◽

Jinzhao Song

Keyword(s):

Point Of Care ◽

Isothermal Amplification ◽

Molecular Test ◽

Rt Pcr ◽

Two Stage ◽

Highly Sensitive ◽

Closed Tube ◽

The Usa ◽

Novel Coronavirus ◽

At Home

<p>The 2019 novel coronavirus (COVID-19) is a newly emerged strain that has never been found in humans before. At present, the laboratory-based reverse transcription-polymerase chain reaction (RT-PCR) is the main method to confirm COVID-19 infection. The intensification of the COVID-19 epidemic overwhelms limited clinical resources in particular, but not only, in developing countries, resulting in many patients not being tested for the infection and in large queues of potentially infected individuals waiting to be tested while providing a breeding ground for the disease. We describe here a rapid, highly sensitive, point-of-care, molecular test amenable for use at home, in the clinic, and at points of entry by minimally trained individuals and with minimal instrumentation. Our test is based on loop mediated isothermal amplification (COVID-19 LAMP) and for higher sensitivity on nested nucleic acid, two stage isothermal amplification (COVID-19 Penn-RAMP). Both tests can be carried out in closed tubes with either fluorescence or colorimetric (e.g., leuco crystal violet LCV) detection. COVID-19 LAMP performs on par with COVID-19 RT-PCR. COVID-19 RAMP has 10 fold better sensitivity than COVID-19 LAMP and COVID-19 RT-PCR when testing purified targets and 100 times better sensitivity than COVID-19 LAMP and COVID-19 RT-PCR when testing rapidly prepared sample mimics. Due to fortunate scarcity of COVID-19 infections in the USA, we were not able to test our assays and methods with patient samples. We hope that such tests will be carried out by colleagues in impacted countries. Our Closed-Tube Penn-RAMP has the potential to significantly reduce false negatives while being amenable to use with minimal instrumentation and training. </p>

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Cryptococcal Meningitis Diagnostics and Screening in the Era of Point-of-Care Laboratory Testing

Journal of Clinical Microbiology ◽

10.1128/jcm.01238-18 ◽

2018 ◽

Vol 57 (1) ◽

Cited By ~ 10

Author(s):

Radha Rajasingham ◽

Rachel M. Wake ◽

Tafese Beyene ◽

Andrew Katende ◽

Emilio Letang ◽

...

Keyword(s):

Cryptococcal Meningitis ◽

Laboratory Testing ◽

Point Of Care ◽

World Health ◽

The Past ◽

Resource Limited ◽

India Ink ◽

Highly Sensitive ◽

The World ◽

Health Organization

ABSTRACT Over the past ten years, standard diagnostics for cryptococcal meningitis in HIV-infected persons have evolved from culture to India ink to detection of cryptococcal antigen (CrAg), with the recent development and distribution of a point-of-care lateral flow assay. This assay is highly sensitive and specific in cerebrospinal fluid (CSF), but is also sensitive in the blood to detect CrAg prior to meningitis symptoms. CrAg screening of HIV-infected persons in the blood prior to development of fulminant meningitis and preemptive treatment for CrAg-positive persons are recommended by the World Health Organization and many national HIV guidelines. Thus, CrAg testing is occurring more widely, especially in resource-limited laboratory settings. CrAg titer predicts meningitis and death and could be used in the future to customize therapy according to burden of infection.

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Towards AI-Enabled Multimodal Diagnostics and Management of COVID-19 and Comorbidities in Resource-Limited Settings

Informatics ◽

10.3390/informatics8040063 ◽

2021 ◽

Vol 8 (4) ◽

pp. 63

Author(s):

Olawande Daramola ◽

Peter Nyasulu ◽

Tivani Mashamba-Thompson ◽

Thomas Moser ◽

Sean Broomhead ◽

...

Keyword(s):

Point Of Care ◽

Low Cost ◽

Personal Data ◽

Disease Diagnosis ◽

Careful Consideration ◽

Diagnostic Methods ◽

Contact Tracing ◽

Patient Treatment ◽

Resource Limited Settings ◽

Resource Limited

A conceptual artificial intelligence (AI)-enabled framework is presented in this study involving triangulation of various diagnostic methods for management of coronavirus disease 2019 (COVID-19) and its associated comorbidities in resource-limited settings (RLS). The proposed AI-enabled framework will afford capabilities to harness low-cost polymerase chain reaction (PCR)-based molecular diagnostics, radiological image-based assessments, and end-user provided information for the detection of COVID-19 cases and management of symptomatic patients. It will support self-data capture, clinical risk stratification, explanation-based intelligent recommendations for patient triage, disease diagnosis, patient treatment, contact tracing, and case management. This will enable communication with end-users in local languages through cheap and accessible means, such as WhatsApp/Telegram, social media, and SMS, with careful consideration of the need for personal data protection. The objective of the AI-enabled framework is to leverage multimodal diagnostics of COVID-19 and associated comorbidities in RLS for the diagnosis and management of COVID-19 cases and general support for pandemic recovery. We intend to test the feasibility of implementing the proposed framework through community engagement in sub-Saharan African (SSA) countries where many people are living with pre-existing comorbidities. A multimodal approach to disease diagnostics enabling access to point-of-care testing is required to reduce fragmentation of essential services across the continuum of COVID-19 care.

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Hepatitis C Virus Infection: A Brief Review

JMS SKIMS ◽

10.33883/jms.v23i1.753 ◽

2020 ◽

Vol 23 (1) ◽

pp. 3-15

Author(s):

Saleem Kamili ◽

Hisham Qadri

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Basic Research ◽

World Health ◽

Hepatitis C Virus Infection ◽

Poor Countries ◽

Diagnostic Assays ◽

Antiviral Therapies ◽

Highly Sensitive ◽

Resource Poor

Hepatitis C, caused by hepatitis C virus (HCV) was originally described as parenterally transmitted non-A non-B hepatitis. Since its discovery in 1989, the field of HCV research has become a shining example of successful translation of basic research wherein in a short of span of just 30 years the virus was discovered, highly sensitive and specific diagnostic assays were developed, epidemiology and clinical characteristics of the disease were well defined and now with the availability of highly efficacious antiviral therapies many countries are already on their way to achieving World Health Organization’s (WHO) elimination targets of hepatitis C by 2030. However, much work needs to be done to eliminate hepatitis C especially in resource poor countries. Most recent data show an estimated 71 million people are currently infected with HCV worldwide and approximately 400,000 people die each year from causes related to HCV. Of these estimates, more than 13 million HCV infected persons are in India and Pakistan (Figure 1). Despite the availability of a cure for hepatitis C, only 20% of those infected patients have been diagnosed (1). In order to achieve the WHO targets of hepatitis C elimination, concerted efforts will have to made to make affordable and reliable diagnostics available worldwide.

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“PHONE-CONNECT” - A Pilot Program Prescribing Phones to Address Digital Health Equity Needs in the COVID-19 Era (Preprint)

10.2196/preprints.23914 ◽

2020 ◽

Author(s):

Gill Kazevman ◽

Marck Mercado ◽

Jennifer Hulme ◽

Andrea Somers

Keyword(s):

Public Health ◽

Health Care ◽

Social Services ◽

Access To Health Care ◽

Health Care Services ◽

Point Of Care ◽

Digital Health ◽

Contact Tracing ◽

Health Care Intervention ◽

Care Intervention

UNSTRUCTURED Vulnerable populations have been identified as having higher infection rates and poorer COVID-19 related outcomes, likely due to their inability to readily access primary care, follow public health directives and adhere to self-isolation guidelines. As a response to the COVID-19 pandemic, many health care services have adopted new digital solutions, relying on phone and internet connectivity. Yet, persons who are digitally inaccessible, such as those struggling with poverty or homelessness, are often unable to utilize these services. In response to this newly highlighted social disparity known as “digital health inequity”, emergency physicians at the University Health Network, Toronto, initiated a program called “PHONE CONNECT”. This novel approach attempts to improve patients’ access to health care, information and social services, as well as improve their ability to adhere to public health directives (social isolation and contact tracing). While similar programs addressing the same emerging issues have been recently described in the media, this is the first time phones are provided as a health care intervention in an emergency department. This innovative ED point-of-care intervention may have a significant impact on improving the health outcomes for vulnerable people during the COVID-19 pandemic, and even beyond it.

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Are social networking platforms the key to effective social distancing for COVID-19? (Preprint)

10.2196/preprints.19722 ◽

2020 ◽

Author(s):

Viknesh Sounderajah ◽

Hutan Ashrafian ◽

Sheraz Markar ◽

Ara Darzi

Keyword(s):

Operating System ◽

Social Networking ◽

Contact Tracing ◽

Text Data ◽

Social Distancing ◽

Web Mapping ◽

Viral Spread ◽

Behavioural Insights ◽

Mapping Software ◽

Incidence And Mortality

UNSTRUCTURED If health systems are to effectively employ social distancing measures to in response to further COVID-19 peaks, they must adopt new behavioural metrics that can supplement traditional downstream measures, such as incidence and mortality. Access to mobile digital innovations may dynamically quantify compliance to social distancing (e.g. web mapping software) as well as establish personalised real-time contact tracing of viral spread (e.g. mobile operating system infrastructure through Google-Apple partnership). In particular, text data from social networking platforms can be mined for unique behavioural insights, such as symptom tracking and perception monitoring. Platforms, such as Twitter, have shown significant promise in tracking communicable pandemics. As such, it is critical that social networking companies collaborate with each other in order to (1) enrich the data that is available for analysis, (2) promote the creation of open access datasets for researchers and (3) cultivate relationships with governments in order to affect positive change.

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Validation of a point-of-care rapid diagnostic test for hepatitis C for use in resource-limited settings

International Health ◽

10.1093/inthealth/ihy101 ◽

2019 ◽

Vol 11 (4) ◽

pp. 314-315

Author(s):

James S Leathers ◽

Maria Belen Pisano ◽

Viviana Re ◽

Gertine van Oord ◽

Amir Sultan ◽

...

Keyword(s):

Point Of Care ◽

Low Cost ◽

Rapid Diagnosis ◽

Direct Acting Antivirals ◽

Resource Limited Settings ◽

Rapid Diagnosis Test ◽

Resource Limited ◽

Specificity And Sensitivity ◽

Hcv Screening ◽

Direct Acting

Abstract Background Treatment of HCV with direct-acting antivirals has enabled the discussion of HCV eradication worldwide. Envisioning this aim requires implementation of mass screening in resource-limited areas, usually constrained by testing costs. Methods We validated a low-cost, rapid diagnosis test (RDT) for HCV in three different continents in 141 individuals. Results The HCV RDT showed 100% specificity and sensitivity across different samples regardless of genotype or viral load (in samples with such information, 90%). Conclusions The HCV test validated in this study can allow for HCV screening in areas of need when properly used.

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