Larsson Gabriella Lillsunde, Karlsson Mats G, Helenius Gisela
Department of Laboratory Medicine, Faculty of Medicine and Health, Örebro University, Örebro - Sweden.
Int J Biol Markers. 2016 May 28;31(2):e218-23. doi: 10.5301/jbm.5000191.
The aim of the study was to investigate whether biobanked liquid-based cytology (LBC) vaginal samples could be reanalyzed for the biomarkers HPV DNA and mRNA without loss of sensitivity.
One hundred LBC samples with ASCUS or CIN1 were tested for HPV DNA and mRNA before and after biobanking. DNA analysis targeted the viral genes E6 and E7, 12 high-risk and 2 low-risk HPV types together with the human control gene HBB, using real-time PCR. The Aptima HPV assay was used for mRNA analysis of 14 high-risk HPV types.
With Aptima there was 84% agreement between results before and after biobanking. The sensitivity and specificity were 0.79 (95% CI, 0.68-0.88) and 0.94 (95% CI, 0.80-0.99), respectively. With the DNA-based method, the agreement between results was 87%, the sensitivity 0.85 (95% CI, 0.75-0.92) and the specificity 0.95 (95% CI, 0.77-1.00). Both methods presented a significant difference between positive results before and after biobanking; McNemar test: p = 0.004, p = 0.003, Cohen's kappa: 0.67 (95% CI, 0.53-0.81), 0.68 (95% CI, 0.52-0.84). Cycle threshold values for the DNA method were higher for all genotypes after biobanking, except for HPV-59. Some loss of sensitivity was seen after biobanking but the concordance between HPV detection before and after biobanking was good for both evaluated methods.
Biobanking of LBC vaginal samples offers a good platform for HPV testing and could be extended to further molecular analyses. However, in order to ensure a valid test result a larger portion needs to be analyzed from the biobanked sample.
本研究的目的是调查生物样本库中的液基细胞学（LBC）阴道样本是否可以在不损失敏感性的情况下重新分析生物标志物人乳头瘤病毒（HPV）DNA和信使核糖核酸（mRNA）。
对100份非典型鳞状细胞意义不明确（ASCUS）或宫颈上皮内瘤变1级（CIN1）的LBC样本在生物样本库保存前后进行HPV DNA和mRNA检测。DNA分析使用实时聚合酶链反应（PCR），针对病毒基因E6和E7、12种高危和2种低危HPV类型以及人类对照基因HBB。Aptima HPV检测用于14种高危HPV类型的mRNA分析。
使用Aptima检测，生物样本库保存前后的结果一致性为84%。敏感性和特异性分别为0.79（95%置信区间，0.68 - 0.88）和0.94（95%置信区间，0.80 - 0.99）。基于DNA的方法，结果一致性为87%，敏感性为0.85（95%置信区间，0.75 - 0.92），特异性为0.95（95%置信区间，0.77 - 1.00）。两种方法在生物样本库保存前后的阳性结果之间均存在显著差异；McNemar检验：p = 0.004，p = 0.003，Cohen's kappa系数：0.67（95%置信区间，0.53 - 0.81），0.68（95%置信区间，0.52 - 0.84）。除HPV-59外，生物样本库保存后DNA方法的所有基因型的循环阈值均较高。生物样本库保存后出现了一定程度的敏感性损失，但对于两种评估方法，生物样本库保存前后HPV检测的一致性都较好。
LBC阴道样本的生物样本库为HPV检测提供了一个良好的平台，并且可以扩展到进一步的分子分析。然而，为了确保有效的检测结果，需要从生物样本库样本中分析更大的部分。
