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In-house reverse transcriptase polymerase chain reaction for detection of SARS-CoV-2 with increased sensitivity.
Kalita, Manash Jyoti; Dutta, Kalpajit; Hazarika, Gautam; Dutta, Ridip; Kalita, Simanta; Das, Partha Pratim; Sarma, Manash P; Banu, Sofia; Idris, Md Ghaznavi; Talukdar, Anjan Jyoti; Dutta, Sangitanjan; Sharma, Ajanta; Medhi, Subhash.
  • Kalita MJ; Laboratory of Molecular Virology and Oncology, Department of Bioengineering and Technology, Gauhati University, Guwahati, Assam, 781014, India.
  • Dutta K; Laboratory of Molecular Virology and Oncology, Department of Bioengineering and Technology, Gauhati University, Guwahati, Assam, 781014, India.
  • Hazarika G; Laboratory of Molecular Virology and Oncology, Department of Bioengineering and Technology, Gauhati University, Guwahati, Assam, 781014, India.
  • Dutta R; Department of Microbiology, GMCH, Guwahati, Assam, 781032, India.
  • Kalita S; Laboratory of Molecular Virology and Oncology, Department of Bioengineering and Technology, Gauhati University, Guwahati, Assam, 781014, India.
  • Das PP; Department of Medicine, GMCH, Guwahati, Assam, 781032, India.
  • Sarma MP; Laboratory of Molecular Virology and Oncology, Department of Bioengineering and Technology, Gauhati University, Guwahati, Assam, 781014, India.
  • Banu S; Department of Medicine, GMCH, Guwahati, Assam, 781032, India.
  • Idris MG; Department of Biotechnology, Assam Down Town University, Guwahati, Assam, 781068, India.
  • Talukdar AJ; Laboratory of Molecular Virology and Oncology, Department of Bioengineering and Technology, Gauhati University, Guwahati, Assam, 781014, India.
  • Dutta S; Laboratory of Molecular Virology and Oncology, Department of Bioengineering and Technology, Gauhati University, Guwahati, Assam, 781014, India.
  • Sharma A; Department of Medicine, GMCH, Guwahati, Assam, 781032, India.
  • Medhi S; Department of Medicine, GMCH, Guwahati, Assam, 781032, India.
Sci Rep ; 11(1): 17878, 2021 09 09.
Article in English | MEDLINE | ID: covidwho-1402125
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.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Reverse Transcriptase Polymerase Chain Reaction / Nucleic Acid Amplification Techniques / COVID-19 Nucleic Acid Testing / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study Limits: Humans Language: English Journal: Sci Rep Year: 2021 Document Type: Article Affiliation country: S41598-021-97502-1

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Reverse Transcriptase Polymerase Chain Reaction / Nucleic Acid Amplification Techniques / COVID-19 Nucleic Acid Testing / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study Limits: Humans Language: English Journal: Sci Rep Year: 2021 Document Type: Article Affiliation country: S41598-021-97502-1