Zappacosta Roberta, Sablone Francesca, Pansa Lucia, Buca Davide, Buca Danilo, Rosini Sandra
Department of Medical, Oral Sciences and Biotechnology, G d'Annunzio University of Chieti-Pescara, Via dei Vestini, 66100 Chieti, Italy.
Surgical Pathology Unit, SS Annunziata Hospital, Via dei Vestini, 66100 Chieti, Italy.
Int J Mol Sci. 2017 Jul 11;18(7):1480. doi: 10.3390/ijms18071480.
Human Papillomavirus (HPV) mRNA test demonstrated high specificity in detecting HPV infections, but studies assessing its efficacy in terms of cancer risk stratification are lacking. Follow-up studies are arduous and expensive. Biobank would be the answer to the problem, although data investigating the effects of long-term storage on RNA preservation are still needed. We addressed these issues by retrieving 202 residual liquid-based cervical specimens, collected from 149 women attending cervical cancer screening during the years 2001-2012. Samples were stored in Adriatic Biobank at room temperature and without any handing. After calculation of RNA yield and purity, mRNA test was retrospectively performed on each samples, to assess analytic and diagnostic performances. Using automated extraction procedures, RNA of good quantity and quality was obtained. The mean value of RNA concentration was 27.5 ng/μL. The mean A260/A280 ratio was 2.1. An invalid mRNA test result was found in 11.9% of the specimens. Neither RNA integrity, nor analytic performances of mRNA test were influenced by the year of sample collection. In total, 62.4% of the specimens tested as mRNA positive; among these, 89.2% were CIN2+. mRNA was detected in all Squamous Cervical Cancer (SCC) cases. Percentage of positive samples increased with the severity of histological diagnosis. mRNA testing, showing specificity and predictive values of 75.6% and 84.4%, respectively, significantly improved the corresponding values for DNA testing. Thus, the reflex mRNA test was demonstrated to be suitable to triage women with persistent cervical lesions. A "one sample for all" approach is possible, with practical benefits for Biobank-based long-term longitudinal studies, diseases prevention, prediction, diagnosis and treatment.
人乳头瘤病毒（HPV）mRNA检测在检测HPV感染方面显示出高特异性，但缺乏评估其在癌症风险分层方面疗效的研究。随访研究艰巨且昂贵。生物样本库可能是解决这一问题的办法，尽管仍需要调查长期储存对RNA保存影响的数据。我们通过检索200份残留的液基宫颈标本解决了这些问题，这些标本采集于2001年至2012年期间参加宫颈癌筛查的149名女性。样本储存在亚得里亚海生物样本库中，室温保存且未作任何处理。在计算RNA产量和纯度后，对每个样本进行回顾性mRNA检测，以评估分析和诊断性能。使用自动化提取程序，获得了数量和质量良好的RNA。RNA浓度的平均值为27.5 ng/μL。平均A260/A280比值为2.1。在11.9%的标本中发现mRNA检测结果无效。样本采集年份既不影响RNA完整性，也不影响mRNA检测的分析性能。总体而言，62.4%的标本mRNA检测呈阳性；其中，89.2%为CIN2+。在所有宫颈鳞状细胞癌（SCC）病例中均检测到mRNA。阳性样本百分比随组织学诊断严重程度增加。mRNA检测的特异性和预测值分别为75.6%和84.4%，显著提高了DNA检测的相应值。因此，反射mRNA检测被证明适用于对患有持续性宫颈病变的女性进行分流。“一个样本用于所有检测”的方法是可行的，这对基于生物样本库的长期纵向研究、疾病预防、预测、诊断和治疗具有实际益处。
