CAJ | 학술논문

目的对2006年广州地区登革热患者的血清进行检测,并对分离的病毒株进行E基因序列分析,以了解可能的传播来源.方法用免疫层析法(ICT)检测患者血清登革病毒IgM和lgG抗体,并用酶联免疫吸附试验(ELISA)检测登革病毒非结构蛋白1(NS1)抗原和IgM抗体;C6/36细胞微量法对发病2 d内患者的血清进行登革病毒的分离培养,并用免疫荧光检测(IFA)及RT-PCR法对病毒分离株进行鉴定;测定分离株DV1-GZ42/06的E基因序列,与国内外参考株、流行株进行同源性比较并构建系统进化树.结果 ICT法检测患者登革病毒IgM和IgG的检出率分别为89.5%(433/484)及38.0%(184/484).ELISA法检测患者血清NSI抗原的检出率为:第1～2天92.7%(38/41)、第3～5天83.3%(70/84)、第6～10天10.9%(5/46);IgM抗体的检出率分别为2.4%(1/41)、51.2%(43/84)及97.8%(45/46).病毒分离培养阳性率为61.0%(25/41),IFA及RT-PCR法汪实为登革1型病毒.分离株Guangzhou/42/06的E基因序列与登革1型病毒标准株Hawaii/45的核苷酸同源件为94.6%;与Thailand/NI09VI04、Vietnam/06和Vietnam/07的同源性高,分别为99.0%、98.6%和98.6%,且处于同一进化分支上,亲缘关系很近,而与广东省2006年前分离株亲缘关系较远.结论登革病毒NS1抗原检测在登革病毒感染的早期诊断中有重要价值.2006年广州地区出现的登革热疫情由输入性病例引起本地暴发流行的可能性大.
목적 대2006년엄주지구등혁열환자적혈청진행검측,병대분리적병독주진행E기인서렬분석,이료해가능적전파래원.방법 용면역층석법(ICT)검측환자혈청등혁병독IgM화lgG항체,병용매련면역흡부시험(ELISA)검측등혁병독비결구단백1(NS1)항원화IgM항체;C6/36세포미량법대발병2 d내환자적혈청진행등혁병독적분리배양,병용면역형광검측(IFA)급RT-PCR법대병독분리주진행감정;측정분리주DV1-GZ42/06적E기인서렬,여국내외삼고주、류행주진행동원성비교병구건계통진화수.결과 ICT법검측환자등혁병독IgM화IgG적검출솔분별위89.5%(433/484)급38.0%(184/484).ELISA법검측환자혈청NSI항원적검출솔위:제1～2천92.7%(38/41)、제3～5천83.3%(70/84)、제6～10천10.9%(5/46);IgM항체적검출솔분별위2.4%(1/41)、51.2%(43/84)급97.8%(45/46).병독분리배양양성솔위61.0%(25/41),IFA급RT-PCR법왕실위등혁1형병독.분리주Guangzhou/42/06적E기인서렬여등혁1형병독표준주Hawaii/45적핵감산동원건위94.6%;여Thailand/NI09VI04、Vietnam/06화Vietnam/07적동원성고,분별위99.0%、98.6%화98.6%,차처우동일진화분지상,친연관계흔근,이여광동성2006년전분리주친연관계교원.결론 등혁병독NS1항원검측재등혁병독감염적조기진단중유중요개치.2006년엄주지구출현적등혁열역정유수입성병례인기본지폭발류행적가능성대.
Objective To detect dengue virus infection by serological method and to determine the sequences of E gene of dengue virus isolated from Guangzhou in 2006.so as to clarify the possible origin of dengue fever.Methods IgM and IgG antibodies to dengue virus were detected by immunochromatographic test(ICT);NSI antigen and IgM antibody were detected by enzyme-linked immunosorbent assay(ELISA).The virus was cultured and isolated from the serum samples within 2 days using C6/36 cell lines and was identified by immuno-fluorescence assay(IFA)and RT-PCR.The E gene of isolated virus DV1-GZ42/06 was sequenced;homological analysis and phylogenetic tree analysis were performed by comparing with the reference strains and epidemic virus strains.Results The positive rates of IgM and IgG of dengue virus in patients were 89.5%(433/484)and 38.0%(184/484)by ICT,respectively.The positive rates of NS1 antigen were 92.7%(38/41)in day 1 to day 2,83.3%(70/84)in day 3 to day 5,and 10.9%(5/46)in day 6 to day 10;and the IgM detection rates were 2.4%(1/41),51.2%(43/84)and 97.8%(45/46)at the same period by ELISA.Twenty-five strains of dengue virus were isolated from 41 serum samples(6 1.O%)and were identified as type 1 dengue virus by IFA and RT-PCR.The sequencing and phylogenetic analysis of the E gene showed that the homology between the isolated Guangzhou/42/06 strain and standard strain Hawaii/45 was 94.6%.and it had a high homology with the Thailand/NI09V104,Vietnam/06.and Vietnam/07 isolates(99.0%,98.6%and 98.6%,respectively)and belonged to the same cladogram,but had low homology with the isolated strain from Guangdong before 2006.Conclusions The detection of NS1 antigen is important in the early diagnosis of dengue fever.The outbreak of dengue fever in Guangzhou in 2006 was possibly caused by the cases from neighboring countries.