Objective To evaluate the detection consistency and power of a multiplex combined real-time PCR detection kits, and provide reference for the prevention and control of influenza plus SARS-CoV-2 infection.
Methods A total of 252 acute respiratory infectious samples were collected in Beijing. Four nucleic acid detection kits (A, B, C for multi nucleic acids and D for nucleic acid of SARS-CoV-2) were used for real time PCR, respectively, to evaluate the consistency of the test results. Three nucleic acid samples of respiratory pathogens in 10-fold dilutions with RNase-free water and 1 nucleic acid samples of SARS-CoV-2 in dilution of 10–1–10–6 folds were used to evaluate the detection power and intra-batch repeatability of kit A.
Results The positive coincidence rates of kit A, B and C to detect influenza A virus, rhinovirus, adenovirus, enterovirus, human metapneumovirus, parainfluenza virus type 3 and 4 and coronavirus NL63 were 100% (all Kappa values=1). The detection power of three assays for influenza B virus, respiratory syncytial virus, parainfluenza virus type 1 and 2, coronavirus OC43, 229E, HKU1 were similar, with the overall consistency rates of 99.17%–100.00% (all Kappa values ≥0.91). The detection power of kit A, B and C were similar (P>0.05). The SARS-CoV-2 negative and positive coincidence rates tested by kit A and kit D were 100%, the intra-batch repeatability showed the coefficient of variatiot (CV) of viral loads tested by these two kits were less than 5% in the dilution range of 10−1–10−3. In the dilution range of 10−4–10−5, the detection power of kit A for open reading frame (ORF)1ab gene was higher (40%) than kit D (30%), as well as for N gene (80% vs. 20%).
Conclusion There were no significant differences in the accuracy of the kit A and other three kits for positive samples with high viral loads, and the detection power were similar. This study provided a multiple choice for large-scale nucleic acid reference for the prevention and control of SARS-CoV-2 infection.
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