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.

Background: The shortage of chemical reagents for severe acute respiratory syndrome - coronavirus 2 (SARS-CoV-2) diagnosis and the surge of SARS-CoV-2 cases, especially in limited-resource settings. Therefore, the combination of an optimal assay kit is necessary. Methods: We compared the ability to screen SARS-CoV-2 among three primer-probe sets added into two different master mixes (Invitrogen SuperScript III One-Step RT-PCR, and LightCycler Multiplex RNA Virus Master). Results: The 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, with R-square and amplification efficiently (AE) were 0.97 and ranged from 107 to 108%, respectively. Conclusions: Our findings indicated TIB-Molbiol, IDT, and Phu Sa primer-probe sets could be beneficial for the laboratory screening of SARS-CoV-2 by Realtime RT-PCR assay of E gene. There is a need for considering the combination of these reagent sets as a new strategy to increase testing capacity for screening programs for COVID-19.