In-house reverse transcriptase polymerase chain reaction for detection of SARS-CoV-2 with increased sensitivity
Abstract As the COVID-19 infection continues to ravage the world, the advent of an efficient as well as the economization of the existing RT-PCR based detection assay essentially can become a blessing in these testing times and significantly help in the management of the pandemic. This study demonst...
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Nature Portfolio
2021
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oai:doaj.org-article:c224515ee7dc42898f7fcfdcc17645482021-12-02T17:19:15ZIn-house reverse transcriptase polymerase chain reaction for detection of SARS-CoV-2 with increased sensitivity10.1038/s41598-021-97502-12045-2322https://doaj.org/article/c224515ee7dc42898f7fcfdcc17645482021-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-97502-1https://doaj.org/toc/2045-2322Abstract As the COVID-19 infection continues to ravage the world, the advent of an efficient as well as the economization of the existing RT-PCR based detection assay essentially can become a blessing in these testing times and significantly help in the management of the pandemic. This study demonstrated an innovative and rapid corroboration of COVID-19 test based on innovative multiplex PCR. An assessment of optimal PCR conditions to simultaneously amplify the SARS-CoV-2 genes E, S and RdRp has been made by fast-conventional and HRM coupled multiplex real-time PCR using the same sets of primers. All variables of practical value were studied by amplifying known target-sequences from ten-fold dilutions of archived positive samples of COVID-19 disease. The multiplexing with newly designed E, S and RdRp primers have shown an efficient amplification of the target region of SARS-CoV-2. A distinct amplification was observed in 37 min using thermal cycler while it took 96 min in HRM coupled real time detection using SYBR green over a wide range of template concentrations. Our findings revealed decent concordance with other commercially available detection kits. This fast HRM coupled multiplex real-time PCR with SYBR green approach offers rapid and sensitive detection of SARS-CoV-2 in a cost-effective manner apart from the added advantage of primer compatibility for use in conventional multiplex PCR. The highly reproducible novel approach can propel extended applicability for developing sustainable commercial product besides providing relief to a resource limited setting.Manash Jyoti KalitaKalpajit DuttaGautam HazarikaRidip DuttaSimanta KalitaPartha Pratim DasManash P. SarmaSofia BanuMd. Ghaznavi IdrisAnjan Jyoti TalukdarSangitanjan DuttaAjanta SharmaSubhash MedhiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021) |
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Medicine R Science Q Manash Jyoti Kalita Kalpajit Dutta Gautam Hazarika Ridip Dutta Simanta Kalita Partha Pratim Das Manash P. Sarma Sofia Banu Md. Ghaznavi Idris Anjan Jyoti Talukdar Sangitanjan Dutta Ajanta Sharma Subhash Medhi In-house reverse transcriptase polymerase chain reaction for detection of SARS-CoV-2 with increased sensitivity |
| description |
Abstract As the COVID-19 infection continues to ravage the world, the advent of an efficient as well as the economization of the existing RT-PCR based detection assay essentially can become a blessing in these testing times and significantly help in the management of the pandemic. This study demonstrated an innovative and rapid corroboration of COVID-19 test based on innovative multiplex PCR. An assessment of optimal PCR conditions to simultaneously amplify the SARS-CoV-2 genes E, S and RdRp has been made by fast-conventional and HRM coupled multiplex real-time PCR using the same sets of primers. All variables of practical value were studied by amplifying known target-sequences from ten-fold dilutions of archived positive samples of COVID-19 disease. The multiplexing with newly designed E, S and RdRp primers have shown an efficient amplification of the target region of SARS-CoV-2. A distinct amplification was observed in 37 min using thermal cycler while it took 96 min in HRM coupled real time detection using SYBR green over a wide range of template concentrations. Our findings revealed decent concordance with other commercially available detection kits. This fast HRM coupled multiplex real-time PCR with SYBR green approach offers rapid and sensitive detection of SARS-CoV-2 in a cost-effective manner apart from the added advantage of primer compatibility for use in conventional multiplex PCR. The highly reproducible novel approach can propel extended applicability for developing sustainable commercial product besides providing relief to a resource limited setting. |
| format |
article |
| author |
Manash Jyoti Kalita Kalpajit Dutta Gautam Hazarika Ridip Dutta Simanta Kalita Partha Pratim Das Manash P. Sarma Sofia Banu Md. Ghaznavi Idris Anjan Jyoti Talukdar Sangitanjan Dutta Ajanta Sharma Subhash Medhi |
| author_facet |
Manash Jyoti Kalita Kalpajit Dutta Gautam Hazarika Ridip Dutta Simanta Kalita Partha Pratim Das Manash P. Sarma Sofia Banu Md. Ghaznavi Idris Anjan Jyoti Talukdar Sangitanjan Dutta Ajanta Sharma Subhash Medhi |
| author_sort |
Manash Jyoti Kalita |
| title |
In-house reverse transcriptase polymerase chain reaction for detection of SARS-CoV-2 with increased sensitivity |
| title_short |
In-house reverse transcriptase polymerase chain reaction for detection of SARS-CoV-2 with increased sensitivity |
| title_full |
In-house reverse transcriptase polymerase chain reaction for detection of SARS-CoV-2 with increased sensitivity |
| title_fullStr |
In-house reverse transcriptase polymerase chain reaction for detection of SARS-CoV-2 with increased sensitivity |
| title_full_unstemmed |
In-house reverse transcriptase polymerase chain reaction for detection of SARS-CoV-2 with increased sensitivity |
| title_sort |
in-house reverse transcriptase polymerase chain reaction for detection of sars-cov-2 with increased sensitivity |
| publisher |
Nature Portfolio |
| publishDate |
2021 |
| url |
https://doaj.org/article/c224515ee7dc42898f7fcfdcc1764548 |
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