L1C-末端共同序列多肽抗体检测宫颈液基细胞学标本中人乳头瘤病毒

L1 C-terminal common sequence polypeptide antibody in detecting human papillomavirus in liquid-based cervical cytology specimens

目的确定由人乳头瘤病毒(HPV)主要外壳蛋白L1 C-末端保守序列多肽诱导的多肽抗体对宫颈脱落细胞内的HPV是否具有良好的检测能力。方法收集宫颈脱落细胞,一式两份,一份采用巢氏PCR检测HPV DNA,另一份以兔抗多肽纯化抗体和小鼠抗多肽抗血清为探针做夹心法ELISA检测标本中的HPV L1,比较两种方法对HPV的检出率差异。用多肽抗体对部分标本做免疫细胞化学检测。结果对收集到的269例宫颈脱落细胞标本用MY09/11引物扩增,检出HPV DNA阳性标本30例,检出率为11.15%;对MY09/11扩增后结果为阴性的标本再用引物GP5+/6+进行扩增。又检出阳性标本51例,检出率为21.34%。将两个PCR检测合计,共检测到81例HPV DNA阳性标本,HPV DNA总检出率为30.11%。对263例脱落细胞标本进行ELISA检测,共检出阳性标本91例,HPV阳性检出率为34.60%,与PCR检测结果相比差异无统计学意义。免疫细胞化学检测结果表明多肽抗体能够特异性地显示有HPV感染的宫颈脱落细胞。结论HPV L1多肽抗体对宫颈脱落细胞内HPV的感染具有良好的检出能力,该抗体在开发用于宫颈癌预防性筛查的HPV检测试剂盒方面具有一定的潜力。

Objective To determine whether the polypeptide antibody induced by human papillomavirus （HPV）L1 C- terminal conserved sequence polypeptide can be used to detect HPV in cervical exfoliated cell specimens. Methods Cervical exfoliated cell specimens were collected and divided into two groups： one was subjected to nested polymerase chain reaction （Nest-PCR） for detection of HPV DNA, and the other was subjected to sandwich enzyme-linked immunosorbent assay （ELISA）, in which rabbit anti-polypeptide purified antibody and mice anti-polypeptide antiserum were used to determine HPV L1 in specimens. The detection rates of HPV were compared between the two groups. Part of the specimens were used for immunocytochemistry detection. Results Nest-PCR with primer MY09/11 detected 30 HPV DNA positive specimens out of a total of 296 ones, with a detection rate of 11.15%. Then the HPV DNA negative specimens were further amplified with primer GP5 ＋/6＋, and 51 （21.34 %） more positive cases were identified, making the total detection rate of Nest-PCR being 30.11 （totally 81 cases）. ELISA identified 91 positive cases out of 263 specimens, with a detection rate of 34.60%, which was not significantly different from that by Nest-PCR. Immunocytochemistry detection results displayed that the polypeptide antibody could specifically reveal cervical exfoliated cells infected with HPV. Conclusion The HPV L1 polypeptide antibody can satisfactorily detect the HPV infection in cervical exfoliated cells, indicating that HPV L1 polypeptide antibody has great potential in developing HPV detection kit for cervical cancer preventive screening.