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目的:应用E2基因缺失的检测了解宫颈病变中HPV16病毒DNA的存在状态,探讨其与宫颈癌的关系。方法:应用多重聚合酶链反应(PCR)对HPV16感染标本将其E2基因的C端、N端、铰链区分别与E6基因同时进行扩增,应用Scion I mage 4.0软件对E2基因、E6基因电泳条带面积灰度值进行半定量分析,判定HPV16是否处于整合状态。结果:在70例宫颈癌标本及42例非癌宫颈病变标本中,HPV16型的感染率分别为67.14%和38.10%。在宫颈癌组中,有40例标本病毒的DNA处于整合状态,6例处于混合状态,1例处于游离状态;在非癌宫颈病变组,有5例病毒DNA处于整合状态,1例处于混合状态,10例处于游离状态。其中感染HPV病毒的正常宫颈组织标本病毒的DNA均处于游离状态。结论:E2基因缺失的检测可以反映HPV16感染状态,对HPV16DNA整合状态的分析可作为判断宫颈病变预后及指导后续治疗的依据。
OBJECTIVE: To detect the presence of HPV16 DNA in cervical lesions by detecting the deletion of E2 gene and to explore its relationship with cervical cancer. Methods: HPV16 infected samples were amplified by polymerase chain reaction (PCR) with C-terminal, N-terminal and hinge regions of E2 gene respectively. The Eion gene was amplified by using Scion I mage 4.0 software. Semi-quantitative analysis of the gray value of the strip area to determine whether the HPV16 is in an integrated state. Results: In 70 cervical cancer specimens and 42 non-cancerous cervical lesions, the infection rates of HPV16 were 67.14% and 38.10% respectively. In the cervical cancer group, DNA of 40 specimens was in the integrated state, 6 cases were in the mixed state and 1 case was in the free state. In the non-cancerous cervical lesion group, 5 cases of the viral DNA were in the integrated state and 1 case was in the mixed state , 10 cases were in free state. The DNA of normal cervical tissue samples infected with HPV virus is in a free state. Conclusion: The detection of E2 gene deletion can reflect the status of HPV16 infection. The analysis of the integrated status of HPV16 DNA can be used as the basis for judging the prognosis of cervical lesions and guiding the follow-up treatment.