CAJ | 학술논문

目的采用自行制备的HPV6b和11型E7蛋白多克隆抗体检测外生殖器尖锐湿疣皮损组织中HPV6b和11型E7蛋白的表达,探讨该抗体在临床检验中的应用价值.方法 55例经临床和病理确诊的尖锐湿疣皮损石蜡标本,用自行制备的HPV6b和11型E7蛋白多克隆抗体进行免疫组化检测,其中18例皮损冰冻标本用RT-PCR法测定HPV6b和11型E7蛋白mRNA表达,分析与免疫组化结果的符合率.结果 55例尖锐湿疣皮损的免疫组化染色显示,HPV6b和11型E7蛋白为胞核染色,且皮损全层表皮细胞均有阳性表达,但基底层阳性细胞较多.HPV6b和11型E7蛋白阳性表达率分别为76.36％(42/55)和58.18％(32/55),总阳性表达率为94.55％(52/55),两蛋白双阳性率为40.00％(22/55),两蛋白均阴性表达为3例(5.45％).18例尖锐湿疣冰冻标本经RT-PCR法检测,HPV6b和11型E7 mRNA阳性表达分别为15例和10例,双阳性表达7例,其阳性表达型别与免疫组化结果完全一致,符合率均为100％.结论该自行制备的HPV6b和11型E7蛋白多克隆抗体免疫组化法检测尖锐湿疣皮损,检测结果直观,可以观察到HPV6b和11型感染细胞在病损中的分布位置.
목적 채용자행제비적HPV6b화11형E7단백다극륭항체검측외생식기첨예습우피손조직중HPV6b화11형E7단백적표체,탐토해항체재림상검험중적응용개치.방법 55례경림상화병리학진적첨예습우피손석사표본,용자행제비적HPV6b화11형E7단백다극륭항체진행면역조화검측,기중18례피손빙동표본용RT-PCR법측정HPV6b화11형E7단백mRNA표체,분석여면역조화결과적부합솔.결과 55례첨예습우피손적면역조화염색현시,HPV6b화11형E7단백위포핵염색,차피손전층표피세포균유양성표체,단기저층양성세포교다.HPV6b화11형E7단백양성표체솔분별위76.36％(42/55)화58.18％(32/55),총양성표체솔위94.55％(52/55),량단백쌍양성솔위40.00％(22/55),량단백균음성표체위3례(5.45％).18례첨예습우빙동표본경RT-PCR법검측,HPV6b화11형E7 mRNA양성표체분별위15례화10례,쌍양성표체7례,기양성표체형별여면역조화결과완전일치,부합솔균위100％.결론 해자행제비적HPV6b화11형E7단백다극륭항체면역조화법검측첨예습우피손,검측결과직관,가이관찰도HPV6b화11형감염세포재병손중적분포위치.
Objective To detect the expressions of human papillomavirus (HPV)-6b/11 E7 proteins in condyloma acuminatum lesions in external genitalia with home-made polyclonal antibodies against HPV-6b/11 E7 proteins,and to estimate the clinical application value of the polyclonal antibodies.Methods Fifty-five tissue specimens were collected from the lesions of patients with clinically and pathologically comfirmed condyloma acuminatum,and subjected to paraffin embedding.An immunohistochemical study was conducted to detect the expressions of HPV-6b/11 E7 proteins in the 55 specimens with home-made polyclonal antibodies against HPV-6b/11 E7 proteins.Reverse transcription (RT)-PCR was performed to quantify the mRNA expressions of HPV-6b/11 E7 in 18 frozen specimens from these patients.The coincidence rate between the two methods was analyzed.Results Immunohistochemical staining showed that HPV-6b/11 E7 proteins were expressed in the nuclei of cells throughout the whole epidermis of the lesions,and especially in the basal layer.Of the 55 specimens,42 (76.36％) stained positive for the HPV-6b E7 protein,32 (58.18％) for the H PV 11 E7 protein,22 (40.00％) for both of them,3 (5.45％) for neither of them,with the positive rate of E7 proteins being 94.55％ (52/55).As RT-PCR showed,the HPV-6b and HPV 11 E7 mRNAs were positive in 15 and 10 out of the 18 frozen specimens,respectively,and both positive in 7 specimens.The HPV types identified by RT-PCR in the 15 specimens were fully consistent with those by the immunohistochemical study.Conclusion The immunohistochemical assay using the home-made anti-HPV-6b/11 E7 polyclonal antibodies can be performed to detect condyloma acuminatum lesions and to observe the distribution of HPV-6b/11-infected cells in an intuitionistic way.