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CC BY-NC-ND 4.0 · J Lab Physicians 2023; 15(03): 383-391
DOI: 10.1055/s-0043-1760752
Original Article

Performance Evaluation of Different RT-PCR Kits for the Direct Detection of SARS-CoV-2 in Preheated Specimens

Rajeev Kumar Jain
1   State Virology Laboratory, Gandhi Medical College, Bhopal, Madhya Pradesh, India
,
Nagaraj Perumal 
1   State Virology Laboratory, Gandhi Medical College, Bhopal, Madhya Pradesh, India
,
Deepti Chaurasia
2   Department of Microbiology, Gandhi Medical College, Bhopal, Madhya Pradesh, India
,
Rakesh Shrivastava
2   Department of Microbiology, Gandhi Medical College, Bhopal, Madhya Pradesh, India
,
Kamlesh Kumar Ahirwar
2   Department of Microbiology, Gandhi Medical College, Bhopal, Madhya Pradesh, India
,
Archa Sharma 
2   Department of Microbiology, Gandhi Medical College, Bhopal, Madhya Pradesh, India
,
Garima Kapoor
2   Department of Microbiology, Gandhi Medical College, Bhopal, Madhya Pradesh, India
,
Jaya Lalwani
2   Department of Microbiology, Gandhi Medical College, Bhopal, Madhya Pradesh, India
› Author Affiliations Funding None.
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Abstract

Background Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has created high demand for molecular kits and consumables for mass screening of suspected individuals. Direct real-time polymerase chain reaction (RT-PCR) assay without nucleic acid extraction has several advantages in saving testing time and cost and helps in the rapid reporting of SARS-CoV-2. The present study evaluated the analytical performance of four SARS-CoV-2 RT-PCR for direct RT-PCR testing using preheated specimens.

Methods A total of 100 clinical specimens were selected and divided into three different groups: (1) group I: 20 SARS-CoV-2 positive specimens with high viral load, viz., low Ct values (< 30 Ct), (2) group II: 50 SARS-CoV-2 positive specimens with low viral load, viz., high Ct values (> 30 Ct), and (3) group III: 30 SARS-CoV-2 negative specimens. Specimens were heat-inactivated at 70°C for 10 minutes and cooled down at 4°C and were evaluated for standard and direct RT-PCR method by using ViralDtect-II Multiplex Real-Time PCR kit, TaqPath COVID-19 Combo kit, COVIDsure Pro Multiplex RT-PCR kit, and Hi-PCR Coronavirus (COVID-19) Multiplex Probe PCR kit.

Results Results showed that except ViralDtect-II kit, the other three TaqPath COVID-19 Combo kit, COVIDsure Pro kit, and Hi-PCR Coronavirus (COVID-19) RT-PCR kit were able to amplify all the SARS-CoV-2 genes in the direct RT-PCR method using preheated specimens. In group I specimens, 100% sensitivity was observed in all three RT-PCR kits. In group II specimens, COVIDsure Pro kit was found to be superior among other kits.

Conclusion Direct RT-PCR method during pandemic situation is valuable and cost effective for the detection of SARS-CoV-2. All three TaqPath COVID-19 Combo kit, COVIDsure Pro kit, and Hi-PCR Coronavirus (COVID-19) RT-PCR kit can be used for direct RT-PCR method and COVIDsure Pro kit performance was found to be superior among all.

Keywords

COVID-19 disease - direct RT-PCR - preheated - RNA extraction - SARS CoV-2


Publication History

Article published online:
30 January 2023

© 2023. The Indian Association of Laboratory Physicians. This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/)

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