scholarly journals A Rapid COVID-19 RT-PCR Detection Assay for Low Resource Settings

Author(s):  
Arunkumar Arumugam ◽  
Matthew L. Faron ◽  
Peter Yu ◽  
Cole Markham ◽  
Season Wong

ABSTRACTQuantitative reverse transcription polymerase chain reaction (RT-qPCR) assay is the gold standard recommended to test for acute SARS-CoV-2 infection. It has been used by the Centers for Disease Control and Prevention (CDC) and several other companies in their Emergency Use Authorization (EUA) assays. RT-qPCR requires expensive equipment such as RNA isolation instruments and real-time PCR thermal cyclers, which are not available in many low resource settings and developing countries. As a pandemic, COVID-19 has quickly spread to the rest of the world. Many underdeveloped and developing counties do not have the means for fast and accurate COVID-19 detection to control this outbreak. Using COVID-19 positive clinical specimens, we demonstrated that RT-PCR assays can be performed in as little as 12 minutes using untreated samples, heat-inactivated samples, or extracted RNA templates. Rapid RT-PCR was achieved using thin-walled PCR tubes and a setup including sous vide immersion heaters/circulators. Our data suggest that rapid RT-PCR can be implemented for sensitive and specific molecular diagnosis of COVID-19 in situations where sophisticated laboratory instruments are not available.

2020 ◽  
Vol 3 (3) ◽  
pp. e29-e34
Author(s):  
Vasileios Bonatsos ◽  
Asif Raza

According to the World Health Organisation there have been 30,055,710 confirmed COVID-19 cases and 933,433 confirmed deaths across 216 countries globally. The availability of the complete SARS-CoV-2 genome relatively early in the epidemic has enabled the development of tests for the diagnosis of COVID-19. There are two broad categories of SARS-CoV-2 diagnostic tests currently in use or development: (1) Real-time reverse transcriptase polymerase chain reaction (RT-PCR) tests and (2) serology tests. RT-PCR is considered the gold standard and preferred method of diagnosis of acute infection. There is, however, a plethora of laboratory-developed and commercial RT-PCR assays with different gene targets. We discuss the value of pre-operative testing for COVID-19 before urological surgery.


Author(s):  
Wanda Markotter ◽  
Jessica Coertse ◽  
Kevin Le Roux ◽  
Joey Peens ◽  
Jacqueline Weyer ◽  
...  

This report describes four suspected rabies cases in domestic dogs that were involved inhuman exposures. In all these cases, the animals were buried for substantial times beforerabies testing was performed. Animal rabies is endemic in South Africa and domestic dogsare the main vector for transmission to humans. Diagnosis of rabies in humans is complicated,and diagnosis in the animal vector can provide circumstantial evidence to support clinicaldiagnosis of rabies in humans. The gold standard diagnostic method, fluorescent antibodytest (FAT), only delivers reliable results when performed on fresh brain material and thereforedecomposed samples are rarely submitted for diagnostic testing. Severely decomposed brainmaterial was tested for the presence of rabies virus genomic material using a quantitativereal-time reverse transcription polymerase chain reaction (q-real-time RT-PCR) assaywhen conventional molecular methods were unsuccessful. This may be a useful tool in theinvestigation of cases where the opportunity to sample the suspected animals post mortem wasforfeited and which would not be possible with conventional testing methodologies becauseof the decomposition of the material.


2021 ◽  
Vol 2 (2) ◽  
Author(s):  
Richard J. Barohn ◽  
Mary Hindle ◽  
Lauren Peck ◽  
Syed Hasan Raza Naqvi

  Since December 2019, COVID 19 pandemic has devastated communities across the world. As number of patients recovered from COVID 19 continue to rise, question of acquired immunity versus chances of re-infection becomes critical to understand the future spread of infection. Here, we present a case of a patient previously recovered from COVID-19, develops new symptoms concerning for possible re-infection with positive reverse transcriptase-polymerase chain reaction (RT-PCR) after few months of initial infection.  


2020 ◽  
Vol 56 (02) ◽  
pp. 087-090
Author(s):  
Saumya Srivastava ◽  
Vidhi Jain ◽  
Vijaya Lakshmi Nag ◽  
Sanjeev Misra ◽  
Kuldeep Singh

AbstractDevelopment of rapid, reliable, and easy diagnostic tests with high-throughput is the need of the hour for laboratories combating the COVID-19 pandemic. While real-time polymerase chain reaction (RT-PCR) is the gold standard for diagnosing active infections, it is expensive and time-consuming. Serological diagnostic assays with a premise to aid rapid contact tracing, immune status determination, and identification of potential convalescent plasma donors hold great promise. Timely diagnosis, effective treatment, and future prevention are key to management of COVID-19.


Author(s):  
Neha Saini ◽  
Prem Pandey ◽  
Mandar Shirolkar ◽  
Atul Kulkarni

Humanity is going through never seen before health crisis due to the outbreak of novel coronavirus or Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2). There are 24.02 million cases and 0.82 million deaths worldwide as of 26th August 2020 due to deadly infection of COVID-19. The disease has been spreading exponentially (R-naught number: 3) and has challenged even the best healthcare infrastructure in the world. With the progression of the disease, the countries shifted the focus from cure to diagnosis and containment to flatten the curve. The review shows that the disease is spreading exponentially while the resources are still limited. We focus upon the probable vectors of the virus, different diagnostic methods with advantages & limitations, and the way forward. This review article covers the different diagnostic methods with more advantages, limitations, and the future sneak-peek into the forthcoming developments for the diagnostic processes such as RT-PCR (Reverse Transcription Polymerase chain reaction).


Author(s):  
Puspa Wardhani ◽  
Trieva Verawaty Butarbutar ◽  
Christophorus Oetama Adiatmaja ◽  
Amarensi Milka Betaubun ◽  
Nur Hamidah ◽  
...  

Background: The diagnostic test for malaria is mostly based on Rapid Diagnostic Test (RDT) and detection by microscopy. Polymerase Chain Reaction (PCR) is also a sensitive detection method that can be considered as a diagnostic tool. The outcome of malaria microscopy detection depends on the examiner's ability and experience. Some RDT has been distributed in Indonesia, which needs to be evaluated for their results. Objective: This study aimed to compare the performance of RightSign RDT and ScreenPlus RDT for detection of Plasmodium in human blood. We used specific real-time polymerase chain reaction abTESTMMalaria qPCRII) and gold standard of microscopy detection method to measure diagnostic efficiency. Methods: Blood specimens were evaluated using RightSign RDT, ScreenPlus RDT, Microscopy detection, and RT-PCR as the protocol described. The differences on specificity (Sp), sensitivity (Sn), positive predictive value (PPV), and negative predictive value (NPV) were analyzed using McNemar and Kruskal Wallis analysis. Results: A total of 105 subjects were recruited. Based on microscopy test, RightSign RDT had sensitivity, Specificity, PPV, NPV, 100%, 98%, 98.2%, 100%, respectively. ScreenPlus showed 100% sensitivity, 98% specificity, 98.2% PPV, 100% NPV. The sensitivity of both RDTs became lower (75%) and the specificity higher (100 %) when using real-time PCR. Both RDTs showed a 100% agreement. RT-PCR detected higher mix infection when compared to microscopy and RDTs. Conclusion: RightSign and ScreenPlus RDT have excellent performance when using microscopy detection as a gold standard. Real-time PCR method can be considered as a confirmation tool for malaria diagnosis.


2021 ◽  
pp. 35-39
Author(s):  
Hanna Sahhar ◽  
Karly Derwitz ◽  
Erica Rubin

Since the declaration of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic in March 2020 by the World Health Organization (WHO), there has been an emergence of a new syndrome termed multisystem inflammatory syndrome in children (MIS-C) associated with COVID-19. MIS-C is defined by the presence of fever, systemic inflammation and multiorgan dysfunction in association with SARS-CoV-2 infection or COVID-19 exposure. Knowledge of this syndrome’s presentation and pathophysiology is constantly evolving as more cases are reported in the literature. This case identifies a 3-month-old patient who tested negative for SARS-CoV-2 antigen, reverse transcriptase polymerase chain reaction (RT-PCR) and antibodies but qualified for MIS-C diagnosis. To the best of our knowledge and through extensive research at the time of diagnosing and reporting this condition to the healthcare authorities, we report the youngest pediatric patient with MIS-C diagnosis. We document this case to contribute to further understanding the variable manifestations of MIS-C and the importance of early diagnosis and treatment with intravenous immunoglobulin (IVIG).


2021 ◽  
Author(s):  
Wan Hang Keith Chiu ◽  
Dmytro Poplavskiy ◽  
Sailong Zhang ◽  
Philip Leong Ho Yu ◽  
Michael D. Kuo

Reverse Transcription-Polymerase Chain Reaction (RT-PCR) is the gold standard for diagnosis of SARS-CoV-2 infection, but requires specialized equipment and reagents and suffers from long turnaround times. While valuable, chest imaging currently only detects COVID-19 pneumonia, but if it can predict actual RT-PCR SARS-CoV-2 status is unknown. Radiogenomics may provide an effective and accurate RT-PCR-based surrogate. We describe a deep learning radiogenomics (DLR) model (RadGen) that predicts a patient's RT-PCR SARS-CoV-2 status solely from their frontal chest radiograph (CXR).


2021 ◽  
Vol 2021 ◽  
pp. 1-4
Author(s):  
Niloufar Bineshfar ◽  
Alireza Mirahmadi ◽  
Fereshteh Karbasian ◽  
Elham Pourbakhtyaran ◽  
Abdollah Karimi ◽  
...  

Coronavirus disease-2019 (COVID-19) which is caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has spread throughout the world causing problems for millions of people. Symptoms of COVID-19 in pediatric patients include both respiratory and gastrointestinal symptoms. The most common symptoms are fever, cough, and fatigue. In this report, we describe a case of a previously well 14-year-old boy, who presented to our emergency department with a complaint of abdominal pain, nausea, and vomiting without fever or respiratory symptoms. He was diagnosed with acute pancreatitis based on an abnormal amylase level and abdomen computed tomography (CT) and later found to be infected by SARS-CoV-2, by a positive reverse transcriptase-polymerase chain reaction (RT-PCR) test.


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