Ardiç Nurittin, Oztürk Oktay, Ergünay Koray, Sezer Ogün
Hipokrat Laboratuvarlari Görüntüleme Merkezi, Istanbul.
Mikrobiyol Bul. 2009 Jul;43(3):463-9.
High-risk human papillomaviruses (HPV) are associated with the development of cervical cancer and its precursor lesions. In addition to cytological screening, nucleic acid testing is the mainstay of diagnosis and follow-up. The molecular tests used for the detection of HPV-DNA in cervical specimens, usually rely on consensus polymerase chain reaction assays that target L1 region of the viral genome. Diagnostic assays that monitor mRNAs from HPV oncogenic proteins, E6 and E7, have also been recently developed. This study was aimed to detect E6/E7 mRNAs from high-risk HPV types 16, 18, 31, 33 and 45 qualitatively by a commercial Nucleic Acid Sequence Based Amplification (NASBA) assay (NucliSENS EasyQ HPV; bioMérieux, France) from cervical specimens. Cervical smear samples were collected from 57 women who had suspected lesions in gynecologic examination and transported by a commercial liquid-based cytology system (ThinPrep Pap Smear Method, Cytyc, USA). Nucleic acid purification was performed by an automated commercial station (NucliSENS easyMAG, bioMérieux, France) as directed by the manufacturer. Presence of viral E6/E7 mRNAs were detected in 38.6% (22/57) of the samples. HPV type 33 mRNA was observed as the most common (11/22, 50%), followed by type 16 (9/22, 41%), 31 (5/22, 22.7%), 45 (4/22, 18.2%) and 18 (1/22, 4.5%). Single and multiple infections with 2 HPV types were identified in 63.6% (14/22) and of 36.4% (8/22) of the positive samples, respectively. The most common co-infection pattern was observed as HPV type 16 and 33 that comprised 13.6% (3/22) of the positive samples. This study was conducted as a preliminary evaluation of commercial NASBA E6/E7 mRNA testing in routine molecular microbiology applications. More studies are required to fully assess the performance of the system for diagnostic laboratories in Turkey.
高危型人乳头瘤病毒（HPV）与宫颈癌及其癌前病变的发生相关。除了细胞学筛查外，核酸检测是诊断和随访的主要手段。用于检测宫颈标本中HPV-DNA的分子检测通常依赖于针对病毒基因组L1区域的共识聚合酶链反应检测。最近也开发了监测HPV致癌蛋白E6和E7 mRNA的诊断检测方法。本研究旨在通过商业核酸序列扩增（NASBA）检测法（NucliSENS EasyQ HPV；法国生物梅里埃公司）对来自宫颈标本的高危HPV 16、18、31、33和45型的E6/E7 mRNA进行定性检测。从57名在妇科检查中疑似有病变的女性中收集宫颈涂片样本，并通过商业液基细胞学系统（美国Cytyc公司的ThinPrep Pap涂片法）进行转运。按照制造商的指示，通过自动商业工作站（法国生物梅里埃公司的NucliSENS easyMAG）进行核酸纯化。在38.6%（22/57）的样本中检测到病毒E6/E7 mRNA。观察到HPV 33型mRNA最为常见（11/22，50%），其次是16型（9/22，41%）、31型（5/22，22.7%）、45型（4/22，18.2%）和18型（1/22，4.5%）。在63.6%（14/22）的阳性样本中鉴定出单一HPV感染，在36.4%（8/22）的阳性样本中鉴定出两种HPV类型的多重感染。最常见的共感染模式是HPV 16和33型，占阳性样本的13.6%（3/22）。本研究是对商业NASBA E6/E7 mRNA检测在常规分子微生物学应用中的初步评估。需要更多研究来全面评估该系统在土耳其诊断实验室中的性能。
