反向杂交法检测HPV基因分型质控品

目的: 反向杂交法检测HPV基因分型质控品。 方法: 采用我室建立的HPV基因分型质控品,以单一型质粒DNA和混合质粒DNA为模板扩增,将扩增好的PCR产物进行导流杂交。 结果: 以单一型质粒DNA为模板扩增时,HPV基因分型结果正确的亚型有18种,该方法的试剂盒对HPV 39,52,53亚型的质粒DNA有漏检。以相同百分比的混合质粒DNA为模板扩增时,试剂盒对HPV 35,39,42,68亚型的质粒DNA有漏检;以不同百分比的混合质粒DNA为模板扩增时,试剂盒对混合质粒中百分比低(5%或者2%)的质粒DNA存在着漏检现象。 结论: 建立的HPV基因分型质控品能够对反向杂交法的试剂盒性能指标进行有效评价,能够满足国内诊断试剂质量监督管理的需要。

Objective: To study the detection of human papiliomavirus genotyping control materials by reverse hybridization assay. Methods: The signal plasmid DNA and mixed plasmids DNA of HPV types as template were amplified.The PCR products were detected by reverse hybridization assay. Results: To use the signal plasmid DNA as template,the genotype of 18 HPV types was accurate.But HPV 39,52 and 53 were not detected.To use the mixed plasmids DNA with the same percentage as template,HPV 35,39,42 and 68 were not detected.To use the mixed plasmids DNA with the different percentage as template,the low percentage plasmid(5% or 2%) was not detected. Conclusion: The constructed human papiliomavirus genotyping control materials effectively evaluate the performance index of genotyping reagent kit with reverse hybridization assay.It will be applied as control material to supervise the quality of diagnostic reagent in our country.