Validation of the performance of a fluorescence quantitative detection system of HCV RNA based on automated extraction

DENG Bo; HE Biao; LIAO Ya-long; HUANG Ge

doi:10.3969/j.issn.1005-4057.2024.01.010

Abstract：Objective To verify the performance indicators of HCV RNA quantitative detection system based on Stream SP96 Automated Nucleic Acid Extractor and ABI 7500 Real-Time PCR System.Methods According to the WS/T 420-2013 Performance Verification Scheme in the Verification of Analytical Performance of Commercial Quantitative Kits by Clinical Laboratory,the Stream SP96 Stream SP96 Automated Nucleic Acid Extractor and ABI 7500 Real-Time PCR System were used to test HCV RNA,and methodological performance verification was performed on the accuracy,precision,linear interval,limit of detection,quantitative limit and anti-interference ability.Results The errors between the mean and target values of the low and high-concentration reference materials were 0.20 and 0.25,respectively,which were less than the logarithmic value of the target value±0.4 log.The intra-batch and inter-batch precision coefficient of variation(CV)values for low-concentration specimens were 0.79%and 1.01%,respectively,and those for high-concentration specimens were 0.52%and 1.22%,respectively,which were all<5%.The linear correlation coefficient r>0.980,and the linear interval was up to 20-1.0×108,showing good linearity(R2=0.997 3).The limit of detection was 20 IU/mL and the lowest limit of quantitation was 50 IU/mL;The interfering substances containing bilirubin(300 mg/L),hemoglobin(300 g/L),and triglyceride(3 000 mg/L)had no effect on the specimen detection results.Conclusion The HCV RNA real-time PCR quantitative testing systems have the accuracy,precision,linear range,limit of detection,limit of quantitation and anti-interference ability in line with the manufacturer's statement,and they can fully meet the clinical requirements for HCV RNA quantitative testing.