Laboratory diagnosis challenges of Covid-19: a Real-time RT-PCR assay-based survey

Authors
Saiedeh Erfanian 1
Danesh Javeshghani 2
Mohsen Farhang 3
Abazar Roustazadeh 4
1 Department of Tissue Engineering, Faculty of Basic Sciences and Advanced Medical Technologies, Royan Institute, ACECR, Tehran, Iran
2 Department of Physiology, Jahrom University of Medical Sciences, Jahrom, Iran
3 Molecular Study and Diagnostic Center, Jahrom University of Medical Sciences, Jahrom, Iran
4 Department of Biochemistry, Department of Advanced Medical Sciences and Technologies, Research Center for Non-communicable Diseases, Jahrom University of Medical Sciences, Jahrom, Iran
10.52547/jmj.19.1.8
Abstract
Introduction: A novel coronavirus has recently been discovered in Wuhan, China in December 2019 named SARS-COV-2. Sensitive molecular assays are now available for detection of SARS-COV-2. The aims of this study were to detect positive subjects of SARS-COV-2 and to survey some challenges that are encountered practically in detection of the virus by Real-time-PCR (RT-PCR) technique.
Materials and Methods: 271 subjects including 105 women and 166 men were registered by census method from 15 April to 15 Jun of 2020. Nasopharyngeal and oropharyngeal swab samples of suspicious cases were collected for RNA extraction to identify SARS-COV-2 by RT-PCR amplification.
Results: 52 women and 82 men were positive for Covid-19. However, technically the results showed that in the time period of 15 April to 15 May of 2020 the contents of received RT - PCR kits were infected with positive control. 8 % of certain negative control samples showed a peak resembling to that of the positive samples in the RT - PCR process. In addition, the results showed that in some of the RT -PCR kits the master mix would produce peaks like positive control without adding the sample to it automatically in 7 % of the tests.
Conclusion: Having different types of negative control for diagnosis of SARS-COV-2 is necessary in extraction and
RT- PCR steps to reduce the chance of having false positive results
Keywords
Coronavirus
Polymerase Chain Reaction
Molecular Study
Quality Control
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