应用H.E染色的鼻咽癌细胞涂片进行DNA含量测定和EB病毒基因原位杂交的研究

分析鼻咽癌常规 H.E染色细胞涂片分别作 Feulgen染色检测 DNA含量和原位杂交检测 EB病毒编码 RNAs(EBERs)的效果 ;将常规 HE染色的 9例正常鼻咽细胞涂片和 38例鼻咽癌细胞涂片用 1%酸性酒精褪色 ,然后作 Feulgen染色 ,应用图象分析仪测定涂片细胞 DNA含量 ,并将 38例鼻咽癌细胞病例的阳性涂片进行 EBERs原位杂交检测 ;结果显示正常鼻咽细胞均为 DNA二倍体核型 ,38例癌阳性涂片中呈 DNA二倍体者 6例 ,DNA非二倍体者 32例 (84.2 % ) ,癌细胞核 EBERs阳性者 35例 (92 .1% ) ;结果表明 HE染色的鼻咽癌细胞学涂片经褪色后作 Feulgen染色和原位杂交检测 ,能较满意进行 DNA和 EBERs分析 ,表明常规 H.E染色和褪色处理均不影响 Feulgen染色和 EB病毒原位杂交的质量效果。本检测方法可靠、可行 ,适用于常规染色细胞学样本的回顾性研究和随访研究 ,并可用于鼻咽癌可疑病人的诊断和鉴别诊断

STUDY OF DNA DETECTION AND IN SITU HYBRIDIZATION FOR EPSTEIN-BARR VIRUS ON H.E STAINED CYTOLOGICAL SMEARS OF NASOPHARYNGEAL CARCINOMA

This study analyzed the suitability and reliability of applying Feulgen stain for DNA analysis and in situ hybridization for Epstein Barr virus(EBV) encoded RNAs(EBERs) on H E stained cytological smears of nasopharyngeal carcinoma(NPC). The HE stained smears, including 9 normal nasopharyngeal smears and 38 NPC ones, were destained using 1% acid alcohol. After that , the DNA ploidy analysis was performed with a CAS 200 image analyzer in all 47 cases and the EBERs was examined on 38 cancer positive smears with in situ hybridization. The normal nasopharyngeal smears showed only diploid DNA pattern. Of the 38 NPC smears, 6 cases showed diploid DNA pattern and 22 cases non diploid pattern(84 2%). EBERs was positive in 35 cases(92 1%). This study demonstrates that the DNA analysis with Feulgen stain and the EBERs detection with in situ hybridization on destained cytological smears with original H E staining is feasible and reliable. This approach could be applied in the retrospective study of the cytological material of patients with long term follow up data and in the diagnosis of suspicious NPC patients.