论文部分内容阅读
AIM:To design and establish a method of multiplex PCRnormalization for simultaneously detecting HBV and HCV.METHODS:Two pairs of primers with a 20 bp jointsequence were used to amplify the target genes of HBVand HCV by two rounds of amplification.After the tworounds of amplification all the products had the jointsequence.Then the joint sequence was used as primersto finish the last amplification.Finally multiplex PCR wasnormalized to a single PCR system to eliminate multiplexfactor interference.Four kinds of nucleic acid extractionmethods were compared and screened.A multiplex PCRnormalization method was established and optimized byorthogonal design of 6 key factors.Then twenty serumsamples were detected to evaluate the validity andauthenticity of this method.RESULTS:The sensitivity,specificity,diagnostic index andefficiency were 83.3%,70%,153.3% and 72.2%,respectively for both HBsAg and anti-HCV positive patients,and were 78.6%,80%,258.6% and 79.2%,respectivelyfor HBsAg positive patients,and were 75%,90%,165%and 83.3%,respectively for anti-HCV positive patients.CONCLUSION:The multiplex PCR normalization methodshows a broad prospect in simultaneous amplification ofmultiple genes of different sources.It is practical,correctand authentic,and can be used to prevent and control HBVand HCV.
AIM: To design and establish a method of multiplex PCR normalization for simultaneously detecting HBV and HCV. METHODS: Two pairs of primers with a 20 bp joint sequence used to amplify the target genes of HBV and HCV by two rounds of amplification. After the tworounds of amplification all the products had the joint sequence. ther the joint sequence was used as primers to finish the last amplification. Four multiplex PCR was normalized to a single PCR system to eliminate multiplexfactor interference. Four kinds of nucleic acid extraction methods were compared and screened. A multiplex PCR normalization method was established and optimized byorthogonal design of 6 key factors. twenty serumsamples were detected to evaluate the validity andauthenticity of this method .RESULTS: The sensitivity, specificity, diagnostic index andefficiency were 83.3%, 70%, 153.3% and 72.2%, respectively for both HBsAg and anti-HCV positive patients, and were 78.6%, 80%, 258.6% and 79.2%, respectivelyfor HBsAg positive patie respectively for anti-HCV positive patients. CONCLUSION: The multiplex PCR normalization methodshows a broad prospect in simultaneous amplification of multiple genes of different sources. It is practical, correct and authentic, and is 75%, 90%, 165% and 83.3% and can be used to prevent and control HBV and HCV.