Bergeron Christine, Cas Frederic, Fagnani Francis, Didailler-Lambert Frederic, Poveda Jean Dominique
Pasteur-Cerba Laboratory, Cergy Pontoise, France.
Acta Cytol. 2006 Jan-Feb;50(1):16-22. doi: 10.1159/000325889.
To validate the utilization of cervical specimens collected in the fixative liquid used in the CYTO-screen System (SEROA, Monaco) for oncogenic human papillomavirus (HPV) DNA detection by the Hybrid Capture II technique (HCII) (Digene, Gaithersburg, Maryland, U.S.A) by reference to cytologic and/or histologic results.
A technical feasibility study was conducted on 3 modalities of sample preparation before HCII technique, 1 with a proteinase digestion, I with PBS washing and I using the Digene sample conversion kit recommended for ThinPrep medium preparation (Cytyc Corp., Boxborough, Massachusetts, U.S.A.). The stability of cells after storage at days 28, 60 and 90 was tested on 26 positive samples (13 with high initial indices and 13 with low initial indices). Results of HPV testing were compared to cytologic and histologic results on a sample of 98 smears already identified as high grade squamous intraepithelial lesion (HSIL) (48) or low grade squamous intraepithelial lesion (LSIL) (50). A retrospective analysis was then performed on 995 HPV tests perfornmed routinely in 2003 in terms of comparison with the corresponding cytologic and/or histologic results.
The HCII technique after direct treatment by proteinase K appeared to be as effective as the Digene sample conversion kit. By using the first technique, all 26 positive cases remained positive at 60 days, but 4 of 13 (30%) with low indices became negative at 90 days. The sensitivity of HPV testing for detecting biopsy- proven cervical intraepithelial neoplasia (CIN) 2 or worse was 100% in the 50 LSIL and 98% in the 48 HSIL samples. In the retrospective study (n = 995), the cytologic diagnoses of atypical squamous cells of undetermined significance (ASC-US) (n=278), LSIL (n = 137) and HSIL (n = 28) were associated with a positive HPVtest in 44%, 75% and 96% of cases, respectively. On a subsample of 156 patients among 278 with a diagnosis of ASC- US, the sensitivity of HPV testingfor detecting CIN 2 or worse was 88%, specificity 57%, positive predictive value 10% and negative predictive value 99%. Performing HPV testing by the HCII technique for cervical specimens collected in the fixative liquid used in the CYTO-screen System is feasible in the context of an ASC-US cytologic diagnosis.
通过参考细胞学和/或组织学结果，验证在CYTO - screen系统（摩纳哥SEROA公司）所用固定液中收集的宫颈标本用于采用杂交捕获II技术（HCII）（美国马里兰州盖瑟斯堡Digene公司）检测致癌性人乳头瘤病毒（HPV）DNA的效用。
在HCII技术之前对3种样本制备方式进行技术可行性研究，一种采用蛋白酶消化，一种采用磷酸盐缓冲盐水（PBS）洗涤，一种使用推荐用于ThinPrep培养基制备的Digene样本转化试剂盒（美国马萨诸塞州博克斯伯勒Cytyc公司）。在26份阳性样本（13份初始指数高和13份初始指数低）上测试了在第28、60和90天储存后细胞的稳定性。将98份已确定为高级别鳞状上皮内病变（HSIL）（48份）或低级别鳞状上皮内病变（LSIL）（50份）涂片样本的HPV检测结果与细胞学和组织学结果进行比较。然后对2003年常规进行的995次HPV检测进行回顾性分析，以与相应的细胞学和/或组织学结果进行比较。
经蛋白酶K直接处理后的HCII技术似乎与Digene样本转化试剂盒一样有效。采用第一种技术时，所有26份阳性病例在60天时仍为阳性，但13份低指数病例中有4份（30%）在90天时变为阴性。在50份LSIL样本和48份HSIL样本中，HPV检测对检测活检证实的宫颈上皮内瘤变（CIN）2级或更严重病变的敏感性分别为100%和98%。在回顾性研究（n = 995）中，意义不明确的非典型鳞状细胞（ASC - US）（n = 278）、LSIL（n = 137）和HSIL（n = 28）的细胞学诊断分别在44%、75%和96%的病例中与HPV检测阳性相关。在278例诊断为ASC - US的患者中的156例亚样本中，HPV检测对检测CIN 2级或更严重病变的敏感性为88%，特异性为57%，阳性预测值为10%，阴性预测值为99%。在ASC - US细胞学诊断的情况下，采用HCII技术对在CYTO - screen系统所用固定液中收集的宫颈标本进行HPV检测是可行的。
