PCR-寡核苷酸微流芯片用于人乳头状瘤病毒分型检测

Human Papilloma Virus typed by PCR based Oligo DNA Microfluid Chip Assay

目的 :应用 PCR-寡核苷酸微流芯片检测方法进行人乳头状瘤病毒 (HPV)分型 ,研究子宫颈上皮内瘤变 (CIN)与 HPV感染型别的关系。方法 :利用简并引物对所有 HPV L1保守区进行扩增 ,再通过寡核苷酸微流芯片上特异性探针对 CIN进行 HPV分型。结果 :41例 CIN病人中 2 2例(5 4 % ) HPV检测为阳性 ,低级 CIN1组高危 HPV感染仅 1 6 % ,CIN2组高危 HPV型感染 5 0 % ,高级 CIN3组高危 HPV型感染 75 % ;与 DNA测序方法比较 PCR-寡核苷酸微流芯片检测方法具有较好的灵敏度。结论 :随着 CIN级别的升高 ,高危 HPV型感染比例增加 ,寡核苷酸微流芯片检测能区分多种 HPV型别 ,可用于临床宫颈上皮内瘤变的 HPV筛查。

Objective: To study the relationship between cervical intraepithelial neoplasia lesions and Human Papilloma Virus (HPV).A PCR based Oligo DNA microfluid chip assay was used for HPV typing.Methods: Consensus primers were used to amplify L1 region of the genome of any HPV.The presence of HPV in a sample is indicated by the formation of amplified DNA.The HPV of CIN lesions was divided by the specific DNA probes on the microfluid chip specific for the amplified region of DNA. Results: Twenty-two out of 41 CIN lesions (54%) were positive for HPV DNA detection.In 'low-grade'CIN 1 group only 16% was associated with the high oncogenic risk HPV types 16,18,or 33,etc.Fifty percent of CIN 2 lesions contained high oncogenic risk HPV types whereas 75% of 'high-grade' CIN 3 lesions contained these HPV types.The Oligo DNA microfluid chip assay had good sensitivity compared with DNA sequencing method.Conclusion: The percentage of high oncogenic risk HPV types increases with the CIN grade elevates.These studies demonstrate that the Oligo DNA microfluid chip assay allows discrimination between most known types of HPV and could be applied to HPV screening of clinical CIN.