HCMV感染标本24小时培养物的原位杂交检测方法  

AN IN SITU HYBRIDIZATION ASSAY FOR DETECTION OF HUMAN CYTOMEGALOVIRUS IN THE 24h CULTURES OF CLINICAL SPECIMENS

作　　者：周越[1] 徐桂林[1] 陆德源[1] 

机构地区：[1]上海第二医科大学微生物学教研室,200025

出　　处：《中华微生物学和免疫学杂志》1995年第3期214-216,共3页Chinese Journal of Microbiology and Immunology

摘　　要：本文探讨了一项检测短期(24 h)培养标本的HCMV感染的原位杂交方法。HCMV核酸探针用HRP直接标记,经原位杂交后与DAB-H_2O_2反应显示紫褐色阳性信号,背景为NFR负染成浅粉红色。经探索,蛋白酶K 50～100μg/ml、NFR负染15 s可取得理想结果。临床上采集22份有异常妊娠史妇女及5份无自然流产史的健康妇女的尿液标本,平行作1天及7天培养,然后同时原位杂交的结果表明,仅1例标本7天检出可疑阳性而1天检出阴性,余检测结果全部吻合,检出阳性率无显著性差异。因此,24h培养标本原位杂交结果可靠,可作为检测HCMV感染的早期快速诊断新方法。An, in situ hybridization assay for detection of HCMV infection in 24 h culture was described. HCMV nucleic acid probe was directly labelled with horseradish peroxidase (HRP),and the positive signal was shown in purple-brown particals in contrast to the light pink background in the nucleus under a light-microscope when the concentration of proteinase K was 50 to l00ug/ml. The urine specimens of 22 women who had abnormal pregnant history were cultured for 1 day and 7 days,and hybridization in situ with the probe was carried out under the same condition simultaneously. The results showed no significant difference. Thus,the results in situ hybridization assay for 24h culture were reliable and themethod could be a new and fastmethod for early diagnosis of HCMV infection.

关 键 词：人类巨细胞病毒 辣根过氧化物酶 原位杂交 

分 类 号：R373[医药卫生—病原生物学]