Cricca Monica, Venturoli Simona, Leo Elisa, Costa Silvano, Musiani Monica, Zerbini Marialuisa
Department of Haematology and Oncological Sciences L e A Seragnoli, Microbiology Section, University of Bologna, Italy.
J Virol Methods. 2009 Jun;158(1-2):180-3. doi: 10.1016/j.jviromet.2009.01.005. Epub 2009 Jan 31.
The presence of HPV 16 E1 and E2 genes was detected in cervical cytological samples using polymerase chain reaction assays. A total of 48 samples were analyzed from patients with HPV 16 infections associated with 13 low-grade cervical intraepithelial neoplasia and 35 high-grade cervical intraepithelial neoplasia. Disruption/deletion sites, within E1 and E2 genes, were detected using 6 primer pairs spanning the entire gene sequences. This technique is not able to recognize mixed DNA forms (integrated plus episomal DNA); therefore, it detects only the presence of pure integrated DNA. Both E1 and E2 genes were detected in 84.6% and in 62.9% of low and high-grade lesions, respectively. The rate of samples with disrupted/deleted genes was significantly higher in high-grade cervical intraepithelial neoplasia than in low-grade cervical intraepithelial neoplasia (P<0.05). In high-grade cervical intraepithelial neoplasia the disruption/deletion pattern involved both E1 and E2 genes and E2 gene was always involved, while in the low grade cervical intraepithelial neoplasia only E1 gene was involved. In conclusion, in high-grade cervical lesions E2 gene seems a suitable target to identify HPV 16 DNA integration into cellular genome.
采用聚合酶链反应检测宫颈细胞学样本中HPV 16 E1和E2基因的存在情况。共分析了48例与13例低级别宫颈上皮内瘤变和35例高级别宫颈上皮内瘤变相关的HPV 16感染患者的样本。使用跨越整个基因序列的6对引物检测E1和E2基因内的破坏/缺失位点。该技术无法识别混合DNA形式(整合型加游离型DNA);因此,它仅检测纯整合型DNA的存在情况。在低级别和高级别病变中,分别有84.6%和62.9%的样本检测到E1和E2基因。高级别宫颈上皮内瘤变中基因破坏/缺失的样本率显著高于低级别宫颈上皮内瘤变(P<0.05)。在高级别宫颈上皮内瘤变中,破坏/缺失模式涉及E1和E2基因,且E2基因总是被涉及,而在低级别宫颈上皮内瘤变中仅涉及E1基因。总之,在高级别宫颈病变中,E2基因似乎是识别HPV 16 DNA整合到细胞基因组中的合适靶点。