Limit of Detection (LOD) of in-house N1N2 CDC real-time RT-PCR assay and commercial kits to detect SARS-CoV-2

Simon Yosonegoro Liem (Clinical Microbiology Laboratory, Microbiology Department, Faculty of Medicine, Universitas Indonesia (FMUI), Jakarta, Indonesia)
Fera Ibrahim (Clinical Microbiology Laboratory, Microbiology Department, Faculty of Medicine, Universitas Indonesia (FMUI), Jakarta, Indonesia)
Andi Yasmon (Clinical Microbiology Laboratory, Microbiology Department, Faculty of Medicine, Universitas Indonesia (FMUI), Jakarta, Indonesia)

Journal of Clinical Microbiology and Infectious Diseases
Vol. 2 No. 2 (2022): Available Online: December 2022

Introduction: There are two types of SARS-CoV-2 real-time RT-PCR (rRT-PCR) kits used in the laboratory in Indonesia, in-house and commercial kits. Our laboratory developed an in-house kit based on N1N2 CDC. In this study, we reported the In-house kit's Limit of Detection (LOD) compared with several commercial kits. Method: This report was an experimental study conducted in Clinical Microbiology Laboratory, Microbiology Department, FMUI in Jakarta. Commercial SARS-CoV-2 RNA (Vircell, Granada, Spain, Lot No. 20MBC137004-R) was used. The LoD was determined using a 2-fold dilution of the RNA in DNase/RNase-free water (Vircell®). The diluted RNA(s) were used as templates for in-house and commercial rRT-PCR kits.  Result: The LOD of in-house rRT-PCR and three commercial kits (BioCoV-19 [Bio Farma], Standard M [SD Biosensor], and Real-Q [BioSewoom]) showed higher sensitivity (3.5 copies/reaction) than Power Chek [Kogenebiotech] (7 copies/reaction).  Conclusion: The LOD of our In-house kit showed high performance in sensitivity and comparable with other commercial kit.