微孔板固相杂交法检测乙型肝炎病毒YMDD变异

目的:建立一种快速、准确、简便的检测乙型肝炎病毒(HBV)YMDD变异的方法。方法:对132份HBVDNA阳性血清标本进行基因测序检测,选取有YMDD变异和无YMDD变异血清标本各20份进行巢式聚合酶链反应,并采用微孔板固相杂交法检测,结果判定阴性和阳性,设直接测序方法为对照进行比较。结果:微孔板固相杂交法检测结果显示,入选的40份标本中19份有变异,21份无变异,其灵敏度为94.74%,特异度为90.48%。结论:微孔板固相杂交法检测乙型肝炎病毒快速、准确、简便,并且可同时检测野生株和变异株类型,尤其适用于大批量标本的检测,便于基层推广。

Detection of YMDD Mutation in Hepatitis B Virus by Coated Microtiter Plate Hybridization

Objective: To establish a rapid, accurate and convenient method for detecting the YMDD mutation of hep-atitis B virus. Methods: One hundred and thirty-two samples with HBV-DNA positive were detected by gene sequencing. Forty samples were recruited in the study.The samples were tested by the Nested-PCR. The product of PCR amplification was tested by coated microtiter plate hybridization to assess the positive and negative results. The results of gene sequence were taken as control test. Results: There were 19 positive and 21 negative results with a sensitivity of 94.74% and a speci-ficity of 90.48% in 40 samples tested by method of coated microtiter plate hybridization. Conclusion: Coated microtiter plate hybridization is a rapid,accurate and sensitive method for detecting the mutation of HBV-DNA,and may be used to replace DNA sequencing and may become a routine method to detect HBV-DNA mutation.