一种HBV DNA 定量检测试剂的准确性分析

目的 评价一种新型HBV DNA 定量检测试剂的准确性。方法 将第3 代HBV DNA WHO 国 际标准品稀释为6 种不同梯度浓度样本，8.5×105、8.5×104、8.5×103、8.5×102、85 及42.5 IU/ml，用待评价 试剂careHBV PCR Assay V3.0 检测，并对实测值和理论值进行相关性分析；以Cobas Taqman HBV V2.0 为参 比试剂，用careHBV PCR Assay V3.0 检测93 份HBV 感染者标本和30 份健康者标本HBV DNA 含量，并对2 种试剂定量检测结果进行相关性和一致性分析；采用巢式PCR 扩增漏检标本的HBV S 区，直接测序法测定 其基因型。结果 careHBV PCR Assay V3.0 检测WHO 国际标准品的实测值和理论值具有线性相关；2 种定 量试剂检测93 份HBV 感染者标本结果均阳性共68 份，检测结果呈线性相关，差异无统计学意义（P >0.05）， 但careHBV PCR Assay V3.0 检测值相对偏低；careHBV PCR Assay V3.0 检测有14 份标本漏检，其中13 份标 本Cobas Taqman HBV V2.0 检测结果位于30 ～ 2 000 IU/ml 间；对1 份漏检标本的基因型检测结果为C 型。 结论 careHBV PCR Assay V3.0 与Cobas Taqman HBV V2.0 试剂检测结果有较好的相关性，但对基因型C 型 的标本可能存在漏检现象。

Accuracy analysis of a type of HBV DNA quantitative detection reagent

Objective To evaluate and analyze the accuracy of a new type of HBV DNA quantitative detection reagent. Methods The 3rd generation HBV DNA WHO international standard was diluted into 6 samples of different concentrations, i.e. 8.5×105, 8.5×104, 8.5×103, 8.5×102, 85 and 42.5 IU/ml. The linear correlations between the theoretical values and the results detected by careHBV PCR Assay V3.0 were analyzed. With Cobas Taqman HBV V2.0 as the reference, HBV DNA content of the specimens from 93 patients infected by HBV and 30 healthy donors were detected by careHBV PCR Assay V3.0 and Cobas Taqman HBV V2.0 simultaneously. And the correlation and consistency of the loads of HBV DNA between the two kinds of results were analyzed. HBV S zone of the samples undetected by careHBV PCR Assay V3.0 was amplified using nested PCR, then the genotypes were determined by direct sequencing. Results The results of the WHO international standard detected by careHBV PCR Assay V3.0 were linearly associated with the theoretical values (R2 = 0.993, P = 0.000). Among the 93 patients infected by HBV, 68 samples were positive by both kinds of quantitative reagents, and the two results were in linear association (R2 = 0.947, P = 0.000); there was no statistical significance between them (t = 1.204, P = 0.231), but the loads of HBV DNA detected by careHBV PCR Assay V3.0 were relatively lower. There were 14 (15.05%) samples undetected by careHBV PCR Assay V3.0, including 13 (46.43%) samples with the results between 30 and 2 000 IU/ml detected by Cobas Taqman HBV V2.0, the genotype of 1 undetected sample was determined to be genotype C. Conclusions The loads of HBV DNA detected by careHBV PCR Assay V3.0 are correlated well with the results detected by Cobas Taqman V2.0. But there may be missed detection of genotype C samples by careHBV PCR Assay V3.0.