CAJ | 학술논문

目的对贵州独山、兴义两地夏秋季不明原因发热病人血清进行登革病毒(DEN)分离及鉴定,从病原学角度证实贵州省人群DEN的感染情况.方法于2005年6至10月份收集贵州独山县、兴义市两地不明原因发热病人血清356份,并接种于生长良好的单层C6/36细胞盲传3代,观察细胞病变,用抗DEN1～4型单克隆抗体通过间接免疫荧光法进行型别鉴定;用DEN NSl基因区特异性通用引物进行RT-PCR检测分离的DEN毒株核酸,经序列测定并作系统发生树分析.结果 3份病人血清标本可使C6/36发生细胞病变,用单克隆抗体、RT-PCR扩增和序列测定,鉴定为DEN2;系统发生树分析证明,分离的病毒与DEN2-43、DEN2-44株系统进化关系最近.结论贵州省独山、兴义两地人群中存在DEN感染.
목적 대귀주독산、흥의량지하추계불명원인발열병인혈청진행등혁병독(DEN)분리급감정,종병원학각도증실귀주성인군DEN적감염정황.방법 우2005년6지10월빈수집귀주독산현、흥의시량지불명원인발열병인혈청356빈,병접충우생장량호적단층C6/36세포맹전3대,관찰세포병변,용항DEN1～4형단극륭항체통과간접면역형광법진행형별감정;용DEN NSl기인구특이성통용인물진행RT-PCR검측분리적DEN독주핵산,경서렬측정병작계통발생수분석.결과 3빈병인혈청표본가사C6/36발생세포병변,용단극륭항체、RT-PCR확증화서렬측정,감정위DEN2;계통발생수분석증명,분리적병독여DEN2-43、DEN2-44주계통진화관계최근.결론 귀주성독산、흥의량지인군중존재DEN감염.
Objective To isolate and identify the Dengue virus(DEN)from the sera of patients with unknown fever in summer and autumn in Dushan and Xingyi areas of Guizhou province,China.Methods From June 2005 to October 2005.356 blood samples were collected from patients with unknown fever in Dushan and Xingyi areas of Guizhou province.The serum samples were inoculated on the C6/36 cell monolayers.After three blind passages,the cytopathie effect(CPE)Was observed.Identification of DEN antigen was earried out by indirect immunofluorescence assay(IFA)with the monoclonal antibody(McAb)against DENl-4 virus.The total RNA was extracted from the serum and tested by RT-PCR with the universal primer for DEN NS region.And determination of the RNA sequenee Was performed,and phylogenetic analysis was carried out.Results Three serum samples caused CPE and were proved as DEN2 positive by IFA,RT-PCR and senquence determination.The phylogenetic tree analysis showed that the isolated virus strains had the closest relations with the systemic evolution of the strains DEN2-43 and DEN2-44.Conclusion DEN infections exist in the population of Dushan and Xingyi of Guizhou,China.