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目的探讨荧光定量聚合酶链(Real-ti me fluorescent quantitative PCR,RT-PCR)方法与传统聚合酶链反应(传统-PCR)方法检测宫颈病变组织DNA中人乳头瘤病毒16型(human papillomavirus,HPVl6)基因的异同。方法 143例(其中维吾尔族标本65例,汉族标本78例)宫颈病变组织样本,均经甲醛固定-石蜡包埋处理。样本分为维、汉2组,对所有样本进行DNA提取,后将DNA样本均分为2份。运用传统-PCR和RT-PCR2种方法检测DNA样本中的HPV16基因,比较其检出率的异同。结果 78例汉族妇女宫颈病变组织DNA样本中,传统-PCR方法和RT-PCR方法对HPV16基因型的检出率分别为51.28%和15.38%,差异有统计学意义(χ2=21.778,P<0.05);65例维吾尔族妇女宫颈病变组织DNA样本中,传统-PCR方法和RT-PCR方法对HPV16基因型的检出率分别为61.54%和18.46%,差异有统计学意义(χ2=19.184,P<0.05)。结论无论是汉族还是维吾尔族,传统-PCR方法对HPV16病毒的检出率均高于RT-PCR方法,因此在临床可以推广简便经济的传统-PCR方法检测高危型HPV16,用于防治宫颈癌。
OBJECTIVE: To detect the expression of human papillomavirus (HPV16) in cervical lesions by Real-ti me fluorescent quantitative PCR (RT-PCR) and traditional polymerase chain reaction ) Gene similarities and differences. Methods Totally 143 cases of cervical lesions (including 65 Uygur specimens and 78 Han specimens) were fixed with formaldehyde and paraffin embedded. Samples were divided into Victoria, Han 2 groups, all samples were DNA extraction, the DNA samples were divided into 2 copies. The HPV16 gene in DNA samples was detected by traditional-PCR and RT-PCR methods, and their similarities and differences were compared. Results The prevalence of HPV16 genotypes in cervical lesions of 78 Han women with cervical lesions was 51.28% and 15.38%, respectively (χ2 = 21.778, P <0.05) ). In 65 Uygur women with cervical lesions, the detection rates of HPV16 genotypes by traditional-PCR and RT-PCR methods were 61.54% and 18.46%, respectively, with significant difference (χ2 = 19.184, P <0.05). Conclusion The detection rate of HPV16 by traditional-PCR method is higher than that of RT-PCR method in both Han and Uygur nationality. Therefore, it is clinically feasible to screen high-risk HPV16 by traditional-PCR method, which is convenient and economical.