Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology
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The severe acute respiratory syndrome coronavirus (SARS-CoV-2) pandemic has resulted in significant shortages of RT-PCR testing supplies including RNA extraction kits. The goal of our study was to determine if a simplified heat-RNA release method would provide comparable detection of SARS-CoV-2 without the need for nucleic acid extraction. ⋯ Five samples that were missed by heat-RNA release method had a mean Ct value: 35 using the automated extraction instrument, indicating a very low viral load. Our findings show that a simple heat-RNA release method is a reasonable alternative for the majority of COVID-19 positive patients and can help overcome the cost and availability issues of RNA extraction reagents.
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Coronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS-COV-2) and represents a global pandemic affecting more than 26 million people and has claimed >870,000 lives worldwide. Diagnostic tests for SARS-COV-2 infection commonly use nasopharyngeal swabs (NPS). As an alternative specimen, we investigated the potential use of the real-time reverse transcriptase PCR (RT-PCR) detection of SARS-COV-2 in saliva samples in large suspected-COVID-19 patients in Kuwait. ⋯ Moreover, the diagnostic sensitivity and specificity of RT-PCR for the diagnosis of COVID-19 in saliva were 83.43 % (95 % CI: 79.07-87.20) and 96.71 % (95 % CI: 94.85-98.04 %), respectively. An analysis of the agreement between the NPS and saliva specimens demonstrated 91.25 % observed agreement (κ coefficient = 0.814, 95 % CI, 0.775-0.854). This study demonstrates that saliva can be a noninvasive specimen for detection of SARS-CoV-2 by RT-PCR.
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Molecular assays based on reverse transcription-loop-mediated isothermal amplification (RT-LAMP) may be useful for rapid diagnosis of the severe acute respiratory syndrome Coronavirus-2 (SARS-CoV-2) because of the easy performance and the option to bypass RNA extraction. ⋯ The variplex RT-LAMP may serve as a rapid test to be combined with a RT-PCR assay to increase the diagnostic accuracy in patients with suspected COVID-19 infection.
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SARS-CoV-2 testing demand has outpaced its supply. Pooling samples for lower risk populations has the potential to accommodate increased demand for SARS-CoV-2 molecular testing. ⋯ We report that 1:4 pooling of samples is specific and associated with an expected 2 CT loss in analytical sensitivity. Instead of running each sample individually, pooling of four samples will allow for a greater throughput and conserve scarce reagents.
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Observational Study
SARS-CoV-2 antibodies, serum inflammatory biomarkers and clinical severity of hospitalized COVID-19 patients.
The involvement of SARS-CoV-2 antibodies in mediating immunopathogenetic events in COVID-19 patients has been suggested. By using several experimental approaches, we investigated the potential association between SARS-CoV-2 IgGs recognizing the spike (S) protein receptor-binding domain (RBD), neutralizing antibodies (NtAb) targeting S, and COVID-19 severity. ⋯ The data presented herein do not support an association between SARS-CoV-2 RBD IgG or NtAb50 levels and COVID-19 severity.