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Comparison of primer-probe sets among different master mixes for laboratory screening of Severe Acute Respiratory Syndrome - Coronavirus 2 (SARS-CoV-2)

Hoang, C. Q.; Hai, H. D.; Hoang, L. T.; Nguyen, A. H.; Nguyen, H. T.; Cao, T. M.; Nguyen, T. T. T.; Phan, L. T.

2020-06-22 emergency medicine
10.1101/2020.06.20.20136242
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BackgroundThere is a shortage of chemical reagents for severe acute respiratory syndrome - coronavirus 2 (SARS-CoV-2) diagnosis and a surge of SARS-CoV-2cases, especially in limited-resource settings. Therefore, the combination of an optimal assay kit is necessary. MethodsWe compared the ability to screen SARS-CoV-2 among three primer-probe sets in two different master mixes, Invitrogen SuperScript III One-Step RT-PCR and LightCycler Multiplex RNA Virus Master. ResultsThe assay with TIB-Molbiol, IDT, and Phu Sa sets for LightCycler Multiplex RNA Virus Master or Invitrogen SuperScript III One-Step RT-PCR showed positive results from a single reaction of triplicate in the three days of 4.8 copies per reaction. R-squared and amplification efficiency were 0.97 and ranged from 107 to 108%, respectively. ConclusionsOur findings indicated that TIB-Molbiol, IDT, and Phu Sa primer-probe sets could be beneficial for the laboratory screening of SARS-CoV-2 by RT-qPCR assay of E gene. There is a need to consider the combination of these reagent sets as a new strategy to increase the testing capacity of screening programs for COVID-19.

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