Investigation of Different Commercially Available Real Time-Polymerase Chain Reaction Kits for SARS-CoV-2 Diagnosis
Introduction: The whole world is facing an ongoing global health emergency of COVID-19 disease caused by the SARS-CoV-2. Real-Time Reverse Transcription-Polymerase Chain Reaction (RT-PCR) is a gold standard in the detection of SARS-CoV-2 infection. Presently, many single tube multiple gene targe...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
JCDR Research and Publications Private Limited
2021-06-01
|
| Series: | Journal of Clinical and Diagnostic Research |
| Subjects: | |
| Online Access: | https://www.jcdr.net/articles/PDF/14977/48726_CE[Ra1]_F[SK]_PF1(AGSK)_redo_(SK)_PFA(KM)_PB(AG_KM)_PN(KM).pdf |
| Summary: | Introduction: The whole world is facing an ongoing global health
emergency of COVID-19 disease caused by the SARS-CoV-2.
Real-Time Reverse Transcription-Polymerase Chain Reaction
(RT-PCR) is a gold standard in the detection of SARS-CoV-2
infection. Presently, many single tube multiple gene target RTPCR kits have been developed and are commercially available
for Corona Virus Disease 2019 (COVID-19) diagnosis.
Aim: To evaluate the performance of seven COVID-19 RT-PCR
kits (DiagSure, Meril, VIRALDTECT II, TruPCR, Q-line, Allplex
and TaqPath) which are commercially available for COVID-19
RT-PCR diagnosis.
Materials and Methods: This observational study was
conducted at the State Virology Laboratory (SVL), Gandhi
Medical College, Bhopal, Madhya Pradesh, India. Seven
commercially available kits have been evaluated on the basis
of: (i) number of SARS-CoV-2 specific gene target; (ii) human
housekeeping genes as internal control; (iii) RT-PCR run time;
and (iv) kit performances to correctly detect SARS-CoV-2
positive and negative RNA samples. A total of 50 RNA samples
(left over RNA) were included, master mix preparation, template
addition and RT-PCR test has been performed according to
kits literature. At the end of PCR run, mean and standard
deviation of obtained cut-off of all kits were calculated using
Microsoft Excel.
Results: All seven RT-PCR kits performed satisfactory
regarding the reproducibility and they could correctly identify
30 positive and 20 negative RNA samples. RNA samples (group
C) having low viral loads with a high Cycle threshold (Ct) value
(>30) were also detected by all these seven kits. Obtained Ct
values of each group was in parallel range in comparison with
the initial testing Ct values. Kits were found to be superior
which contains primers and probes for three SARS-CoV-2
specific gene targets, have human housekeeping gene as
internal control and taking less time to complete RT-PCR.
Conclusion: All seven COVID-19 RT-PCR kits included in this
study demonstrated satisfactory performance and can be used
for the routine molecular diagnosis of COVID-19 disease. |
|---|---|
| ISSN: | 2249-782X 0973-709X |