人乳头瘤病毒16和18型核酸检测用质控品制备及其应用

Preparation and application of internal quality control material for nucleic acid testing of human papillomavirus type 16 and 18

目的制备可模拟真实临床标本的HPV-16和HPV-18核酸检测用质控品,并用于上海地区临床实验室的室间质量评价。方法分别选择整合有HPV-16和HPV-18 L1基因的宫颈癌细胞系(Siha和Hela)进行培养,收集细胞后用HPV国际标准品定值并分装,对其均匀性和稳定性进行评价,然后将样本盘发送至参评实验室,对回报结果进行分析评价。结果均匀性评价结果显示质控品间均匀性良好,稳定性评价结果表明质控品检测结果随时间延长无明显趋势变化。参评实验室对高浓度样本的检测符合率较高,低浓度样本的检测符合率较低。对于不分型项目,26%参评实验室由于上报假阴性结果导致出错。对于分型项目,不同检测方法的结果间没有显著差异,样本检测出错主要是因为错检或漏检某一亚型所致。结论制备的核酸检测用质控品的均匀性和稳定性良好,可有效用于临床实验室室间质量评价。质评结果显示上海地区参评实验室HPV核酸检测质量整体较好,个别实验室检测能力尚需提高。

Objective To prepare the internal quality control material simulating clinical specimens for nucleic acid testing of human papillomavirus（ HPV） type 16 and 18 and evaluate the performance in the detection and genotyping of HPV type 16 and 18 by the external quality assessment（ EQA） of laboratories in Shanghai. Methods The cultured Siha and Hela cervical carcinoma cell lines known to be integrated with L1 gene of HPV-16 and HPV-18 were collected as internal quality controls. The international units of HPV nucleic acid in the cells were determined by tracing the international standards and dispensed into EP tubes. The homogeneity and stability of the prepared control material were evaluated. The panels were distributed to the EQA participants in Shanghai area and the return results were summarized and evaluated. Results The homogeneity evaluation for the prepared control material showed acceptable results that there were no significant differences among the different laboratories. The stability evaluation indicated that the no significant linear trend was found with extended test time. The EQA results showed that high coincidence rate was achieved for the samples with high concentration of HPV. Among the laboratoried which could not identify the genotype,26% reported wrong results due to false-negative data. For HPV genotyping,there was no significant difference for the results between the laboratories with different detection methods. The false and missed results of detection for certain HPV subtype led to unqualified reports. Conclusion The acceptable homogeneity and stability of the prepared quality control material for nucleic acid testing may effectively evaluate the internal quality of the laboratories in HPV DNA testing. The results in this srudy indicated that most of the participants in Shanghai showed good performance in detecting HPV DNA,but a few laboratories have to improve their operation.