Physical state of human papillomavirus type 16 in cervical intraepithelial lesions and cancers determined by two different quantitative real-time PCR methods.
Physical state of human papillomavirus type 16 in cervical intraepithelial lesions and cancers determined by two different quantitative real-time PCR methods.
复制标题
通过两种不同的实时定量 PCR 方法测定宫颈上皮内病变和癌症中人乳头瘤病毒 16 型的物理状态。
作者：
S. Szostek;B. Biesaga;B. Zawilińska;M. Klimek;M. Kosz
本研究的目的是分析用于评估人乳头瘤病毒（HPV16）载量和病毒基因组状态的新型多重qPCR方法与参考qPCR方法之间的相关性。该研究是在100个含有癌前病变和癌的HPV16阳性样本中进行的。采用两种PCR方法检测hpv16e2和E6基因载量。针对RNase P开放阅读框进行qPCR，将E2和E6的负载归一化为细胞数。病毒基因组的物理状态以每个细胞E2/E6拷贝数的比率来确定。在分析的100份样本中，多重qPCR和qPCR检测E2和E6病毒载量差异无统计学意义，相关系数分别为0.98和0.97。多重qPCR检测到病毒基因组完整状态的样品占19%，混合状态的样品占73%，独立状态的样品占8%，qPCR检测到病毒基因组完整状态的样品分别为17%，79%,4%。多重qPCR估计的综合和单项形式的患病率高于qPCR获得的患病率（ch2, p < 0.0001），但在恶性前和恶性诊断的样本中，无论使用哪种方法，都没有发现显著差异。与qPCR相比，多重qPCR的敏感性为93.7%，特异性为100%，阳性预测值为100%。综上所述，多重qPCR检测HPV16的载量和病毒基因组状态的频率是一种敏感和特异性的参考方法。在一个反应管中同时检测E2和E6基因降低了检测成本。
The aim of this study was to analyse the correlation between a new multiplex qPCR assay and a reference qPCR assay for assessment of the human papillomavirus (HPV16) load and the viral genome status. The study was performed on 100 HPV16 positive samples containing premalignant lesions and carcinomas. HPV16 E2 and E6 gene loads were assessed by two PCR methods. The load of E2 and E6 was normalized to the cell number by qPCR targeting the RNase P open reading frame. The physical state of the viral genome was determined as a ratio of E2/E6 copies number per cell. Among 100 samples analysed, there were no statistically significant differences in the E2 and E6 viral load evaluated by multiplex qPCR and qPCR, the correlation coefficients were 0.98 and 0.97, respectively. There were 19% of samples with the integrated, 73% with mixed and 8% with episomal state of viral genome detected by multiplex qPCR and 17%, 79%, 4%, respectively, found by qPCR. Prevalence of integrated and episomal forms estimated by multiplex qPCR was higher than the one obtained by qPCR (Chi2, p < 0.0001), but in samples with premalignant and malignant diagnoses no significant differences were demonstrated regardless of the methods used. Sensitivity and specificity of multiplex qPCR were 93.7% and 100% as compared with qPCR, the positive predictive value was 100%. In summary, the multiplex qPCR assay in respect of HPV16 load and the frequency of viral genome status was shown to be a sensitive and specific reference method. Simultaneous estimation of E2 and E6 genes in one reaction tube reduces the cost of testing.
影响因子：
5.5
作者：
Moscicki, Anna-Barbara;Schiffman, Mark;Villa, Luisa L.
通讯作者：
Villa, Luisa L.
影响因子：
56.9
作者：
DYSON, N;HOWLEY, PM;HARLOW, E
通讯作者：
HARLOW, E