宫颈癌石蜡标本中人乳头状瘤病毒检测方法的初步探讨

Detection of human papilloma virus (HPV) in cervical cancer from paraflin-embedded samples.

目的:探讨适合检测宫颈癌石蜡包埋标本中高危型HPV的方法。方法:用高危型HPV-DNA HC2检测及PCR扩增联合DNA测序法,对25例宫颈癌(宫颈鳞癌23例,宫颈腺癌2例)进行高危型HPV检测。每例取石蜡包埋组织块中的癌组织,21例术前曾于门诊取材(新鲜脱落细胞)行高危型HPV-DNA HC2检洲。结果:(1)21例宫颈癌石蜡标本, PCR扩增法高危型HPV阳性检出率为66．7%;HC2高危型检测阳性检出率为47．6%;(2) PCR扩增法灵敏度为66．7%,特异度为33．3%,与参照标准的符合率为61．9%,阳性预测值为85．7%,阴性预测值为14．3%;(3)HC2检测法灵敏度为44．4%,特异度为33．3%,与参照标准的符合率为42．9%,阳性预测值为80．0%,阴性预测值为9．1%;(4)石蜡标本保存时间与HPV检出率有关。半年以内标本HPV检出率为92．3%(12/13),半年以上者为33．3% (4/12),两者差异有统计学意义(P=0．004)。结论:HPV-DNA扩增(PCR)联合测序法是目前回顾性研究宫颈癌石蜡包埋标本中HPV感染的可行方法;石蜡标本保存时间与HPV检出率有关,石蜡包埋时间越短、标本越新鲜,检出率越高。

Objective： To compare two methods for detection of high-risk HPV in cervicad cancer in paraffin-embedded samples from cervical cancer. Methods：Cervical cancer tissues from paraffin-embedded samples of each case were collected. DNA specimens, extracted from paraffin-embedded tissues of 25 patients with cervical cancer （ including 23 squamous cell cacinomas and 2 adenocarcinomas, were analysed for detection of high-risk HPV by two methods：HPV-DNA HC-2 and PCR combined with sequencing of HPV-DNA. In addition, fresh brush-off cells from 21 patients were detected by high-risk HPV-DNA HC-2 before the operations. Results： （1）Using the PCR combined with sequencing and HC-2 to detect 21 cases, high-risk HPV was detected in 66. 7% and 47.6%, respectively; （2） The sensitivity of PCR was 66. 7% ,the specificity was 33.3%, the coincidence rate was 61.9%, the positive predicted value was 85.7% ,the negative predicted value was 14.3% ; （3）The sensitivity of HC-2 was 44.4% ,the spocificity was 33.3%, the coincidence rate was 42.9%, the positive predicted value was 80.0%, the negative predicted value was 9.1% ; （4） The positive rate of high-risk HPV detection was related to the kept time of the paraffin-embedded specimens. The specimens stored within half a year accounted for 52% （ 13/25 ） and 48% specimens were stored more than half a year. The positive rates of high-risk HPV detection in the two groups were 92.3 % （ 12/13 ） and 33.3% （4/12）, respectively （ P = 0.004）. Conclusion： HPV-DNA PER combined with sequencing is a feasible method to detect high-risk HPV in paraffin-embedded tissues from cervical cancer. The positive rate of HPV detection is associated with the kept time of the paraffin-embedded specimens： the shorter the kept time, the fresher of the specimen, the higher the positive rate of HPV detection was.