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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

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

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【作者】 李莉宋武琦李小光马培林张凤民

【Author】 LI Li,SONG Wu qi,LI Xiao guang, et al . (Department of Ophthalmology,the First Clinical Medical College of Harbin Medical University,Harbin 150001,China)

【机构】 哈尔滨医科大学第一临床医学院眼科哈尔滨医科大学微生物学教研室哈尔滨医科大学微生物学教研室 黑龙江哈尔滨150001黑龙江哈尔滨150086黑龙江哈尔滨150086

【摘要】 目的 分析2 0 0 3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,Ad V)感染情况及其型别。方法 采集5 9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- RFL P)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% ( 4 1/5 9)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,5 3.6 % ( 2 2 / 4 1)的CPE阳性标本可以扩增出腺病毒核酸,RFL P分析证实6 8.18% ( 15 / 2 2 )的腺病毒感染为Ad3型,31.82 % ( 7/ 2 2 )的腺病毒感染为Ad7。结论2 0 0 3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- RFL P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

【Abstract】 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.

【关键词】 急性结膜炎腺病毒细胞培养PCR-RFLP
【Key words】 ConjunctivitisAdenovirusViral isolationPCR-RFLP
【基金】 哈尔滨市青年科学研究基金项目 (编号 :2 0 0 3AFQXJ0 4 0 )
  • 【文献出处】 中国微生态学杂志 ,Chinese Journal of Microecology , 编辑部邮箱 ,2005年02期
  • 【分类号】R777.31
  • 【被引频次】9
  • 【下载频次】175
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