病毒分离和PCR-RFLP法对急性结膜炎标本中腺病毒感染及其型别的分析

目的分析2003年下半年收集的哈尔滨医科大学第一临床医学院急性结膜炎标本中的腺病毒(Adenovirus，AdV)感染情况及其型别。方法采集59例临床确诊为急性结膜炎患者的结膜拭子标本，在观察和分析接种细胞的病变(CPE)情况的基础上，采用聚合酶链式反应-限制性片段长度多态性分析(PCR—RFLP)法，特异性检测CPE阳性细胞中腺病毒核酸的存在情况，并对PCR阳性扩增产物进行病毒型别的分析。结果70．7％(41／59)的急性结膜炎标本接种于培养细胞后可以引起明显的CPE；其中，53．6％(22／41)的CPE阳性标本可以扩增出腺病毒核酸，RFLP分析证实68．18％(15／22)的腺病毒感染为Ad3型，31．82％(7／22)的腺病毒感染为Ad7。结论2003年下半年哈尔滨市急性结膜炎主要以腺病毒3、7型感染为主。提示结合病毒分离培养和PCR—RFLP方法，可以广泛应用于急性结膜炎相关的病毒感染及其型别的进一步分析与研究。

Analysis of Adenovirus infection related with acute conjunctivitis by means of viral isolation and PCR-RFLP

Objective To analyze the Adenovirus infection related with acute conjunctivitis occurred from June to December 2003 in Harbin city,China.Methods Viral isolation was carried out through the examination of cyto pathogenic effect (CPE) on the Hela cell culture inoculated separately with 59 samples of conjunctive scra pings from patiens with acute conjunctivitis.Then Adenovirus nucleic acid were detected by polymerase chain reaction (PCR) and the viral typing were analyzed by restriction fragment length polymorphism (RFLP).Results After 48 hours incubation,70 7% (41/59) of the cell culture with samples inoculation were showed significant coty pathogenic effect formation.Then 53 6% (22/41) of CPE showed cell samples were amplified as Adenovirus DNA positive by PCR method and the Adenovirus were further classified to be 68 18% (15/22) of Ad 3 type and 31 82% (7/22) of Ad 7 type by RFLP analysis.Conclusions Those results were certificated that the occurrence of acute conjunctivitis was caused mainly with Adenovirus,especially with Ad 3 and Ad 7 types.It also suggested it combination of viral isolation and PCR RFLP method would be used to analyze the viral pathogen related with acute conjunctivitis in the future.