Evaluate The Application Of Different Pcr Technologies In Detecting HBV-DNA

Objective Evaluate the application of domestic and Cobas Ampli Prep/Cobas Taq Man 48 PCR technology in detecting HBV-DNA.Method Serum samples were collected from 190 patients with chronic B hepatitis who were treated in the First Affiliated Hospital of Anhui Medical University. All the samples were tested with domestic reagent and Cobas Taq Man reagent. The performance of the two reagents were compared and analyzed by correlation analysis and Kappa test.Results1. For the total 190 specimens, 72 specimens which below the detection offline of domestic fluorescence quantitative PCR reagent(HBV-DNA≤500IU/ml) and 7specimens which exceeded the upper limit of Roche Cobas Taq Man fluorescence quantitative PCR reagent(HBV-DNA≥1.7×108IU/ml) should be excluded. The rest111 specimens were compared by correlation analysis, and the results were statistically significant(P < 0.05, R2= 0.6707). It showed that domestic fluorescence quantitative PCR reagents and roche Cobas Taq Man reagent presented a linear correlation.2. The total 111 specimens were divided into HBe Ag(+) group and HBe Ag(-) group.The correlation between two groups was analyzed, and the difference was statistically significant(P < 0.05). The results showed that domestic fluorescence quantitative PCR reagents and roche Cobas Taq Man reagent of two groups presented a linear correlation; It also showed that the HBe Ag(+) group(R2=0.5864) had a higher correlation than that of HBe Ag(-) group(R2=0.5226)3. The total 190 domestic reagent specimens could be divided into four groups,according to the guideline of different HBe Ag status on antiviral treatment of HBV DNA in the 2010 version of the prevention and control of chronic hepatitis B. They were HBe Ag(+), HBV DNA < 20000 IU/ml group, HBe Ag(+), HBV DNA >20000 IU/ml group, HBe Ag(-), HBV DNA < 20, 00 IU/ml group and HBe Ag(-)and HBV DNA > 20, 00 IU/ml group. The four groups were compared by correlation analysis. The results indicated that, for the patients with a high viral load,domestic reagent test was more reliability than the Roche COBAS reagent which was not influenced by HBe Ag status; however, domestic reagent were unreliable when tested the specimens from low viral load patients, especially from the HBe Ag(-) groups;4. All the specimens for consistency analyzed according to the different HBe Ag status and DNA load for antiviral requirement. In HBe Ag(+) group there were 98 cases with the HBV load was higher or equal to 20000 IU/ml(HBV DNA≥20000IU/ml).This group was set to test positive. While the HBV load which below 20000 IU/ml(HBV DNA < 20000 IU/ml) was set to test negative; The result showed that domestic reagent had a highly consistent(Kappa=0.752) with Cobas Taq Man reagent testing; Basing on the requirements of antiviral, there were 92 cases in HBe Ag(-) group. The HBV load which higher or equal to 2000IU/ml( HBV DNA≥20,00IU/ml) was set to test positive, and the HBV load which below2000IU/m l(HBV DNA < 2000 IU/ml) was set to test negative; The result suggested that the consistency between domestic reagent for detection and Cobas Taq Man reagent testing(Kappa=0.381) is poor.Conclusion1. The two test methods had poor conformity in HBe Ag-negative group.2. Cobas Taq Man reagent ought to be used to quantitate HBV-DNA in patients with HBe Ag-negative chronic type-B hepatitis; however, domestic reagent could be used in HBe Ag positive group.