实时荧光聚合酶链反应技术与第2代杂交捕获法检测高危亚型人乳头瘤病毒的比较

Application of real-time polymerase chain reaction and hybrid capture Ⅱ in high-risk human papillomavirus detection

目的比较实时荧光聚合酶链反应技术(实时PCR)检测人乳头瘤病毒(HPV)16、18型与第二代杂交捕获法(HC-Ⅱ)检测13种HPV高危亚型。方法采用实时PCR与HC-Ⅱ分别检测宫颈脱落细胞标本中的HPV16、18型和13种HPV高危亚型,并进行测序分析。结果HPV16、18型实时PCR总的检出率为24%,13种HPV高危亚型HC-Ⅱ总的检出率为46.6%;高度鳞状上皮细胞内病变(HSIL)组实时PCR和HC-Ⅱ的检出率均较低度鳞状上皮细胞内病变(LSIL)组要高,分别为54%与21%、84.3%与65.6%。HPV16、18型在HPV高危亚型中的总检出率为51%。有2例宫颈炎组标本HC-Ⅱ检测阴性而实时PCR检测阳性,测序结果与实时PCR一致。结论实时PCR法检测HPV16、18较HC-Ⅱ灵敏度高,但检测的HPV亚型少,相比之下HC-Ⅱ更适合于临床上HPV高危亚型的筛查检测。

Objective To compare real time polymerase chain reaction （real time PCR） with hybrid capture Ⅱ （HC -Ⅱ ） for detection of genotype 16,18 and 13 high risk genotypes of human papillomavirus （HPV）. Methods Real time PCR and HC -Ⅱ were used to detect genotype 16,18 and 13 genotypes of high risk HPV respectively. The results were confirmed with DNA sequencing. Results The total detection rate of real time PCR of genotypes 16,18 and HC-Ⅱ of 13 HPV high risk genotypes was 24% and 46. 6% respectively. The detection rates of the two methods for high-grade squamous intraepithelial lesion （HSIL） group were both higher than that of low-grade squamous intraepithelial lesion （LSIL） ,which was 54% vs 21% ,84.3% vs 65.6%. The overall detection rate of HPV genotype 16,18 in the 13 genotypes was 51%. And DNA sequencing suggested that the two cervicitis samples results were the same with real lime PCR results,which were HPV positiveS,not negative. Conclusion Real time PCR is more sensitive method for detection of HPV 16,18 genotypes than HC-Ⅱ ,but the detected quantity of genotypes is few. HC-Ⅱ is more suit able to detection of high-risk type HPV infection in cervix.