【摘要】 目的分析2种DNA提取方法对人乳头状瘤病毒(HPV)分型杂交结果的影响,为进一步提高临床检验质量和改进检测技术提供理论依据。方法收集326例宫颈脱落细胞标本,分别采用磁珠法和常规法提取HPV核酸,应用核酸分子快速导流杂交技术进行21种HPV亚型检测,研究不同的方法对HPV-DNA浓度、纯度和分型结果的影响。结果 1常规方法抽提的DNA纯度、浓度分别为(1.64±0.09)μg/m L和(88±9)μg/m L;磁珠法抽提的DNA纯度、浓度分别为(1.79±0.06)μg/m L和(105±11)μg/m L;磁珠法所抽提的DNA纯度和含量分别高于常规法(P<0.01)。2常规法检测出现聚合酶链反应(PCR)抑制物的例数为6例,而磁珠法则为0例。3常规法和和磁珠法HPV分型的总阳性率、单一感染率、多重感染率分别为12.9%、12.0%、0.9%和12.0%、1.2%、13.2%,两法一致性检验Kappa=0.907,结果具有较好的一致性。结论磁珠法可获得质量较好的HPV DNA,分型结果满意,可以有效去除PCR反应抑制物,操作简便,节省时间,更容易实现大批量样本检测,适合临床检验应用。更多还原

【Abstract】 Objective To investigate the influence of different extractive methods of DNA on the detection of genotypes of human papillomavirus(HPV). Methods The 326 cervical epithelium samples were treated by magnetic beads method and traditional method;The samples were checked for 21 types HPV by flow through hybridization and gene chip technology,The different extractive methods were used to study the efficiency of HPV DNA extraction and genoty-ping. Results 1 DNA purification and concentration by traditional extractive method were(1.64 ±0.09) μg/m L and(88 ±9) μg/m L,while that used of HPV-DNA by magnetic beads extractive method were(1.79 ±0.06) μg/m L and(105±11)μg/m L,markedly increased compared with those used by traditional method(P<0.01). 2 Polymerase chain reaction(PCR)inhibitors were detected from 6 samples by the traditional extractive method,but no PCR reaction inhibitor was detected from the samples by magnetic beads extractive method. 3 The rates of positive,single types of infectionand multiple infection in the HPV types treated by the traditional method and magnetic beads method were 12.9%,12.0%,0.9% and 12.0%, 1.2%, 13.2%, respectively. The value of Kappa was 0.907, showing a high degree of consistency. Conclusion The magnetic beads method is suitable for detection of HPV-DNA from cervical epithelium samples,that is easier to get rid of the PCR reaction inhibitors and bath processing samlples,which ensures the quality of HPV DNA,It is applicable in clinical laboratory.更多还原