二种分子杂交技术定量检测临床标本中HBV DNA的比较

Comparison of two hybridization quantitative assays for detecting HBV DNA

目的 第 2代核酸杂交捕获系统 (HCⅡ )与分枝链DNA信号放大系统 (bDNA)定量检测HBVDNA的临床应用比较。方法 4 0例乙型肝炎病毒表面抗原阳性的慢性乙型肝炎患者临床血清标本共 79份 ,采用 2种杂交方法定量检测HBVDNA。结果 HCⅡ与bDNA定量检测HBVDNA的阴阳性符合率为 85 .9% ,相关系数r =0 .96 ,P <0 .0 0 1;经敏感性比较 ,HCⅡ灵敏度较bDNA提高 10 1拷贝 /ml～ 10 2 拷贝 /ml。HCⅡ与bDNA定量检测HBVDNA结果的换算方程为 :HCⅡ (HBVDNA拷贝 /ml) =1.0 14×〔bDNA (HBVDNAEq/ml)〕0 .963 2 ;或bDNA (HBVDNAEq/ml) =4 .0 34×〔HCⅡ (HBVDNA拷贝 /ml)〕0 .9574。在操作上HCⅡ比bDNA更为简便、快速。结论 HCⅡ与bDNA法检测HBVDNA具有较好的相关性 ,可用于临床HBVDNA水平的定量检测、监控及抗病毒药物的筛选、考核。

Objective To compare two different hybridization assays for quantitative measurement of HBV DNA. Methods 79 clinical serum samples from 40 HBsAg positive patients were selected. The HBV DNA of these samples were measured using a solution hybridization assay based on a RNA/DNA hybrid capture (HCⅡ TM ) system (Digene) and a signal amplification assay based on branched DNA (bDNA) technology (Bayer). Results In comparison with the HCⅡ and bDNA assays, concordant results were found in 73 ( 85.9% ) samples, and the results of the two assays were closely correlated ( r = 0.96 , P < 0.001 ). Compared with bDNA, the sensitivity of HCⅡ was slightly higher by about 10 1～10 2 copies/ml. The conversion formula for the two assays results was: HBV DNA by HCⅡ (copies/ml)= 1.014 ×〔HBV DNA by bDNA (equivalent/ml, Eq/ml)〕 0.9632 ; or: HBV DNA by bDNA (Eq/ml)= 4.034 ×〔HBV DNA by HCⅡ (copies/ml)〕 0.9574 . The protocol for the HCⅡ was less complex and required shorter time (4 h) than that of bDNA (24 h). Conclusions The specificity and sensitivity of Digene HC Ⅱ system were almost the same as that of Bayer bDNA assay in the quantitative measurement of serum HBA DNA. Thus, this study suggested that HCⅡ also can be recommended for monitoring HBV DNA levels during antiviral treatment.