多通道实时荧光定量PCR检测HPVDNA结果分析

目的探讨多通道实时荧光定量PCR检测HPVDNA的效果，并应用于日常检测工作。方法对临床1100例宫颈分泌物标本采用多通道实时荧光定量PCR仪进行8种高危HPVDNA分型及定量检测。8种高危HPV型别为主要高危型HPV16、18、45、31和次要高危型HPV33、52、58、67。结果在1100例标本中排除了14例B球蛋白为阴性的标本，剩余1086例标本总体的内标平均值为3．71×10^6IU／ml。随机重复性试验和对每个型别的各浓度标本进行的重复性试验每次扩增均为阳性，型别符合率为100％。共检测出阳性标本287例，HPV16、HPV18／45、HPV31、次要高危型和合并感染各有46、17、14、146和64例，分别占总阳性标本的16％、6％、5％、51％和22％。所有标本绝对定量值的分布近似于正态分布。结论多通道荧光定量PCR检测HPVDNA灵敏度高、复性好，可用于临床HPV感染的筛查与宫颈病变程度预测以及患者术后疗效观察。

Detection of HPV DNA by using multi-channel fluorescence real time quantitative PCR assay

Objective To evaluate the detection of human papilloma virus （HPV） DNA by using multi-channel fluorescence real time quantitative PCR assay. Methods Eight genotypes of high risk HPV （HPV16, 18, 45, 31 and HPV33, 52, 58, 67） DNA were quantitatively detected by using multi-channel fluorescence Real-Time PCR in 1 100 samples of cervical secretions. Results In 1 100 samples, 14 were excluded because of negativeβ-globin; while in 1 086 included samples, 13-globin was assayed simultaneously for absolute quantization of genomic DNA with an average level of 3.71E ＋6 IU/ml. All positive results were obtained both in random repeatability test and repeatability test for each types with various concentrations, with a type coincidence rate of 100%. HPV DNA was detected in 287 out of 1 086 samples. The genotype distributions were HPV16 （ 16% ）, HPV18/45 （6%）, HPV31 （5%）, HPV33/52/58/67 （51%） and 64 samples were detected as co-infected HPV （ 22% ）. The absolute quantitative value distribution presented as normal distribution. Condusiou The multi-channel Real-Time fluorescence quantitative PCR assay is sensitive andrepeatable in detection of HPV DNA, which can be used for detection of HPV infection in clinical evaluation of cervical lesions.