Pooled RNA sample reverse transcriptase real time PCR assay for SARS CoV-2 infection : A reliable, faster and economical method
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.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2020 |
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| Erschienen: |
2020 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:15 |
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| Enthalten in: |
PloS one - 15(2020), 7 vom: 31., Seite e0236859 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Gupta, Ekta [VerfasserIn] |
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| Links: |
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| Themen: |
Coronavirus Envelope Proteins |
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| Anmerkungen: |
Date Completed 21.08.2020 Date Revised 18.12.2020 published: Electronic-eCollection Citation Status MEDLINE |
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| doi: |
10.1371/journal.pone.0236859 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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NLM313059268 |
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| 100 | 1 | |a Gupta, Ekta |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Pooled RNA sample reverse transcriptase real time PCR assay for SARS CoV-2 infection |b A reliable, faster and economical method |
| 264 | 1 | |c 2020 | |
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| 500 | |a Date Completed 21.08.2020 | ||
| 500 | |a Date Revised 18.12.2020 | ||
| 500 | |a published: Electronic-eCollection | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a 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 | ||
| 520 | |a 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 | ||
| 520 | |a 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 | ||
| 520 | |a 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 | ||
| 520 | |a 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 | ||
| 650 | 4 | |a Evaluation Study | |
| 650 | 4 | |a Journal Article | |
| 650 | 7 | |a Coronavirus Envelope Proteins |2 NLM | |
| 650 | 7 | |a RNA, Viral |2 NLM | |
| 650 | 7 | |a Viral Envelope Proteins |2 NLM | |
| 650 | 7 | |a envelope protein, SARS-CoV-2 |2 NLM | |
| 700 | 1 | |a Padhi, Abhishek |e verfasserin |4 aut | |
| 700 | 1 | |a Khodare, Arvind |e verfasserin |4 aut | |
| 700 | 1 | |a Agarwal, Reshu |e verfasserin |4 aut | |
| 700 | 1 | |a Ramachandran, Krithiga |e verfasserin |4 aut | |
| 700 | 1 | |a Mehta, Vibha |e verfasserin |4 aut | |
| 700 | 1 | |a Kilikdar, Mousumi |e verfasserin |4 aut | |
| 700 | 1 | |a Dubey, Shantanu |e verfasserin |4 aut | |
| 700 | 1 | |a Kumar, Guresh |e verfasserin |4 aut | |
| 700 | 1 | |a Sarin, Shiv Kumar |e verfasserin |4 aut | |
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