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

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Simon Yosonegoro Liem https://orcid.org/0000-0002-7464-1797
Fera Ibrahim https://orcid.org/0000-0002-3732-0812
Andi Yasmon https://orcid.org/0000-0002-3999-4976

Keywords

Limit of Detection, LOD, SARS-CoV-2, Coronavirus

Abstract

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.

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