Research Of Rapid Detection Of Hepatitis B Virus And Hepatitis C Virus

Objective A loop-mediated isothermal amplification (LAMP) method hadbeen developed to detect hepatitis C virus (HCV) and different subtypes ofhepatitis B virus (HBV) rapidly.Methods LAMP primers were designed according to the specific DNAsequences of HBV B, C and D genotypes, respectively. LAMP reactions werecarried out with optimal conditions. The specificity of LAMP primer sets werevalidated among HBV subtypes of B, C and D. The specificity and sensitivity ofLAMP compared with Real time PCR by assaying clinical samples of HBV. Beside,LAMP primers were designed according to the specific DNA sequences of HCVgenome, LAMP reactions were carried out with optimal conditions. The specificityof LAMP primer sets were validated by HCV genome. The specificity andsensitivity of LAMP compared with the mothod of clinical used by assayingclinical samples of HCV.Results LAMP method could detect HBV B, C and D subtype effectively intens of minutes and the detection limits of HBV-B subtype were17copies, HBV-Csubtype25copies and HBV-D subtype10copies. Compared with Real-time PCR,the sensitivity of LAMP enhances about1000times. Also LAMP method coulddetect HCV genome effectively in two hours and detection limits of HCV genomewere180copes/mol.Conclusion LAMP could detect HCV genome and different HBV genotypesrapidly with high sensitivities and specificities, which should be applied to clinicallaboratory.