一种荧光PCR方法用于高危型HPV基因分型检测

目的: 建立一种可以检测高危型人乳头瘤病毒(HR-HPV)的荧光PCR方法,为临床宫颈癌的筛查提供新的检测试剂盒。 方法: 选择高危型HPV的L1区基因序列作为靶序列,应用Primer Premier 5.0软件在选取的序列区域设计引物和探针,建立和优化该检测方法。通过评价其准确性、灵敏度和特异性,建立一种荧光PCR方法,用于高危型HPV基因分型的检测。 结果: 用HPV L1基因型质控品进行检测时,该试剂盒针对高危型HPV L1基因质控品均具有较好的阳性结果;检测灵敏度约为10~100 拷贝·反应体系^-1;低危型HPV L1基因质控品则为阴性结果,用正常人白细胞全基因组DNA进行检测时,没有HPV特异性扩增信号出现。 结论: 建立的高危型HPV基因分型荧光PCR检测方法具有良好的准确性、检测灵敏度和特异性,可以用于临床宫颈癌的筛查。

Objective: To establish a fluorescence PCR method for high risk HPV genotyping test which can provide a novel test reagent kit for cervical cancer screening. Methods: The high risk HPV L1 gene sequence was selected as the target sequence,and the primers and probes were designed in the selected sequence region with Primer Primer 5.0 software.Then the fluorescence PCR method for HR-HPV genotyping test was established and optimized.Finally,the fluorescence PCR method was established after evaluating accuracy,sensitivity and specificity. Results: Using this reagent kit to test the HPV L1 genotyping control materials,HR-HPV types showed better positive signals but LR-HPV types showed negative signals.The test sensitivity of fluorescence PCR detection was about 10~100 copies reaction^-1 for multiple HR-HPV detection.And there were no specific HPV amplification signals for human leukocyte genomic DNA. Conclusion: The Fluorescence PCR method was proved to be an effective HPV detection method with good accuracy,sensitivity and specificity which could be widely applied to cervical cancer screening.

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