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目的探讨应用基因芯片检测宫颈石蜡组织标本中人乳头状瘤病毒(HPV)感染的可能性及其临床意义。方法收集解放军总医院诊断为宫颈鳞状上皮病变的石蜡组织标本40例,其中宫颈浸润性鳞癌18例,宫颈上皮内瘤变(CIN)Ⅲ12例,CINⅠ4例,CINⅡ6例。从组织中提取DNA后采用基因芯片检测23种常见HPV基因亚型,即PCR扩增后产物在基因芯片上进行杂交。同时选用10例经基因芯片检测16型和18型基因阳性的宫颈鳞癌的石蜡组织切片做原位杂交。基因芯片检测结果与部分原位杂交结果进行比较并分析。结果基因芯片检测的18例宫颈鳞癌HPV高危亚型均为阳性(100%),其中1例为混合阳性;12例CINⅢ中11例为高危亚型阳性(91·7%),1例阴性;6例CINⅡ的宫颈病变中高危型5例阳性,低危型1例阳性;4例CINⅠ中有2例低危型阳性、2例阴性;宫颈鳞癌和CINⅢ组与CINⅠ和Ⅱ组比较,差异有统计学意义(U=80·0,P<0·01)。10例宫颈鳞癌基因芯片HPV16型和18型阳性组织中,原位杂交同型探针6例检测显示阳性。结论HPV基因芯片技术可用于检测多种亚型,特异性强,敏感性高,对HPV感染亚型的鉴别及宫颈癌的预防和治疗具有重要意义。
Objective To investigate the possibility and clinical significance of detecting human papillomavirus (HPV) infection in cervical paraffin tissue samples by using gene chip. Methods Forty paraffin-embedded specimens of cervical squamous epithelial lesions were collected from PLA General Hospital. Among them, 18 were squamous cell carcinoma of the cervix, 12 were cervical intraepithelial neoplasia (CIN) Ⅲ, 4 were CIN Ⅰ and 6 were CIN Ⅱ. After DNA was extracted from tissues, 23 common HPV genotypes were detected by gene chip, that is, the products after PCR amplification were hybridized on the gene chip. In the meantime, 10 cases of cervical squamous cell carcinoma positive for type 16 and 18 gene by gene chip were selected for in situ hybridization. Gene chip test results and partial in situ hybridization results were compared and analyzed. Results HPV DNA was found to be positive (100%) in all 18 cases of cervical squamous cell carcinoma detected by gene chip, of which 1 was mixed positive. Among the 12 cases of CIN Ⅲ, 11 were high risk subtypes (91.7%) and 1 negative ; 6 cases of CIN Ⅱ cervical lesions in 5 cases of high-risk type and low-risk type 1 positive; 4 cases of CIN Ⅰ in 2 cases of low-risk type positive and 2 cases negative; cervical squamous cell carcinoma and CIN Ⅲ group compared with CIN Ⅰ and Ⅱ group, The difference was statistically significant (U = 80 · 0, P <0.01). In 10 cases of cervical squamous cell carcinoma gene microarray HPV16 and 18 positive tissues, 6 cases of in situ hybridization probes showed positive results. Conclusion HPV gene chip technology can be used to detect a variety of subtypes, strong specificity, high sensitivity, identification of HPV subtypes and cervical cancer prevention and treatment of great significance.