Preliminary Application Of High-throughput Sequencing To Detect HBV Multi-drug Resistance Sites And Genotyping

Objective:To verify the preliminary application value of high-throughput sequencing technology in detecting HBV multi-drug resistance sites and genotyping,and to explore whether high-throughput sequencing methods can detect HBV resistance sites and genes can provide guidance for clinical treatment.Methods:Sixty-five patients with HBV multi-resistance sites and genotypes were screened by PCR reverse dot hybridization in the Department of Infectious Diseases,Jiangsu Provincial People’s Hospital.High-throughput sequencing techniques were used to retest the specimens.The results of the two detection methods were compared to analyze the differences and to evaluate the application value of high-throughput sequencing in the detection of multiple drug resistance sites and genotypes of HBV.Results:1.The results of high-throughput sequencing technology: The results of the genotypes of 65 patients examined were: 9 cases of type B,54 cases of type C,2cases of type B+C,and mainly type C,accounting for 83.08%.Of the 65 patients,37 had no mutations detected,and 28 patients had detected mutations(25 in type C,2 in type B,and 1 in type B+C).The detection rate was 43.08%.Among the 28 positive mutant patients,14 resistant mutation sites were detected,including 169 T,173L,180 M,181T,181 V,184A,184 I,184L,184 S,204I,204 V,207I,250 I and 250 L.2.The results of PCR reverse dot hybridization showed that the results ofgenotyping in 65 patients were 9 cases of type B,53 cases of type C,and 1 case of type B+C.No genotypes were detected in 2 cases,and C types were mainly detected.,accounting for 81.54%.Of the 65 patients,38 had no resistance mutations,and 27 had resistant mutations(22 in C type,4 in B,and B+C).The detection rate was 41.54%.Among 27 mutation-positive patients,11 mutation sites were detected,including180 H,180M,181 T,181V,184 L,204I,204 V,236T,250 I,250L,and 250 V.3.High-throughput sequencing provides specific quantification of genotypes and resistance sites,and is superior to PCR reverse dot hybridization that provides qualitative results;high-throughput sequencing can detect >5% or more of the mutant strains,while PCR The point-to-point hybridization technique can only be detected when the mutation rate is >35%,and some resistance sites with a mutation rate of less than 35% are missed.Conclusions:High-throughput sequencing can provide quantitative results for HBV resistance genotypes and mutation sites,and intuitively analyze drug resistance.High-throughput sequencing can detect mutation sites when the mutation rate is less than 5%,and requires low mutation frequency.It can measure more sites than the PCR-reverse-point hybridization method and has important significance for the discovery of potential drug resistance sites.It has practical clinical application value and can monitor potential drug resistance sites during treatment of patients.Dynamic changes are conducive to discovering and monitoring new drug-resistant forms,providing basis for drug selection and changing treatment plans for patients.