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PLoS One ; 15(7): e0236859, 2020.
Article in English | MEDLINE | ID: mdl-32730368

ABSTRACT

BACKGROUND: Corona virus disease 2019 (COVID-19) which initially started as a cluster of pneumonia cases in the Wuhan city of China has now become a full-blown pandemic. Timely diagnosis of COVID-19 is the key in containing the pandemic and breaking the chain of transmission. In low- and middle-income countries availability of testing kits has become the major bottleneck in testing. Novel methods like pooling of samples are the need of the hour. OBJECTIVE: We undertook this study to evaluate a novel protocol of pooling of RNA samples/elutes in performance of PCR for SARS CoV-2 virus. STUDY DESIGN: Extracted RNA samples were randomly placed in pools of 8 on a 96 well plate. Both individual RNA (ID) and pooled RNA RT-qPCR for the screening E gene were done in the same plate and the positivity for the E gene was seen. RESULTS: The present study demonstrated that pool testing with RNA samples can easily detect even up to a single positive sample with Ct value as high as 38. The present study also showed that the results of pool testing is not affected by number of positive samples in a pool. CONCLUSION: Pooling of RNA samples can reduce the time and expense, and can help expand diagnostic capabilities, especially during constrained supply of reagents and PCR kits for the diagnosis of SARS-CoV-2 infection.


Subject(s)
Betacoronavirus/genetics , Coronavirus Infections/diagnosis , Coronavirus Infections/epidemiology , Pneumonia, Viral/diagnosis , Pneumonia, Viral/epidemiology , RNA, Viral/genetics , Real-Time Polymerase Chain Reaction/methods , Reverse Transcriptase Polymerase Chain Reaction/methods , COVID-19 , Coronavirus Envelope Proteins , Coronavirus Infections/virology , Diagnostic Tests, Routine/economics , Diagnostic Tests, Routine/methods , Humans , India/epidemiology , Mass Screening/economics , Mass Screening/methods , Pandemics , Pneumonia, Viral/virology , Prospective Studies , RNA, Viral/isolation & purification , Real-Time Polymerase Chain Reaction/economics , Reproducibility of Results , Reverse Transcriptase Polymerase Chain Reaction/economics , SARS-CoV-2 , Viral Envelope Proteins/genetics
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