2003年广州市登革1型病毒流行株的分离及NS2基因序列分析  

Isolation and Sequence Analysis of Dengue Virus Type 1 from Suspected Dengue Virus Infected Patients in Guangzhou in 2003

作　　者：卢业成[1] 江振友[2] 陈万山[1] 任瑞文[3] 方美玉[3] 王少珍[1] 程刚锋[3] 田小东[3] 刘建伟[3] 尹炽标[1] 

机构地区：[1]广州市第八人民医院,广州510060 [2]暨南大学医学院病原生物学教研室,广州510630 [3]广州军区联勤部军事医学研究所微生物室,广州510507

出　　处：《热带医学杂志》2004年第6期666-669,共4页Journal of Tropical Medicine

摘　　要：目的对广州市区2003年仲夏一批疑似登革病毒感染的患者进行确诊,并从基因水平分析流行株的可能来源。方法用免疫层析法(ICT)检测早期疑似患者的DV-IgM和IgG抗体;同时用逆转录-聚合酶链反应(RT-PCR)、细胞培养病毒分离分别进行病原的分离和鉴定;对所分离到的毒株进行基因克隆、测序,并与国际参考株及国内流行株相应的序列进行同源性分析,结合患者的流行病学资料推测本次流行株的可能来源。结果发病5d内患者DV-IgM抗体阳性率为56.5%(13/23),发病5～10d的患者DV-IgM抗体阳性率为66.7%(16/24);DV-IgG抗体无1例阳性。从18份发病5d内患者的血标本中分离病毒7份,经RT-PCR和基因测序检测证实为DV1感染;用RT-PCR检测30份早期患者血标本,患者的阳性率为83.3%(25/30)。基因序列分析表明,2003年流行株与登革1型病毒柬埔寨流行株及我国1997、1999年登革1型病毒流行株的同源性最高,分别为97%、97%、98%。所有患者在发病前两个月均在广州市区某大院内居住,无输血史、无外出史。结论2003年广州登革热流行为登革1型病毒感染所致,推测广东可能存在登革1型病毒的疫源地。Objective To identify the dengue virus isolated from Guangzhou,Guangdong province in 2003 and to discuss the possible origin.Methods Laboratory diagnosis involved three testing methodologies:virus isolation by cell culture,a reverse transcription polymerase chain reaction (RT PCR)assay,and serologic assays using ICT technique. Results 13/23 (56.5%)were positive in IgM ICT test in 5 day; 14/24 (66.7%)were positive in IgM ICT test in 5 to 10 day; none were positive in IgG ICT test.Virus were isolated from seven of 18 serum samples of patients in 5 day and were identified as dengue virus type 1 by RT PCR and DNA sequence analysis. 25/30(83.3%)showed positive in RT PCR assay. DNA sequence of the isolated virus showed 97%,97%and 98%homology with those of dengue virus type 1 strains of dengue hemorrhagic fever outbreak in Cambodia,in China 1997 and 1999 respectively. All patients live in Guangzhou and have no recent travel history outside the area in two months before infection.Conclusion The isolated virus from Guangzhou,Guangdong province in 2003 is belong to dengue virus type 1, and there may be a plague spot in Guangdong.

关 键 词：登革病毒 病毒分离 逆转录-聚合酶链反应 序列分析 

分 类 号：R373.33[医药卫生—病原生物学]