中华流行病学杂志
中華流行病學雜誌
중화류행병학잡지
CHINESE JOURNAL OF EPIDEMIOLOGY
2012年
8期
768-773
,共6页
张如胜%欧新华%宋克云%袁洁%陈田木%肖姗%孙边成
張如勝%歐新華%宋剋雲%袁潔%陳田木%肖姍%孫邊成
장여성%구신화%송극운%원길%진전목%초산%손변성
环境%禽流感病毒%职业暴露%H5N1抗体
環境%禽流感病毒%職業暴露%H5N1抗體
배경%금류감병독%직업폭로%H5N1항체
Environment%Avian influenza virus%Occupational exposure%H5N1 antibody
目的 对长沙市家禽市场职业暴露人群进行禽流感病毒(AIV)H5N1亚型抗体水平和环境AIV核酸检测,并对环境中AIV H5N1亚型的血凝素(HA)基因进行测序分析.方法 抽取长沙市1个区和1个县,各选择2个城区或乡镇家禽市场进行职业暴露人群H5N1抗体和环境AIV核酸检测.利用单放射免疫扩散溶血实验(SRH)对102份家禽市场职业暴露人员血清标本进行H5N1抗体检测,real-time PCR方法检测160份家禽市场环境标本(污水、禽类粪便、羽毛和禽类笼具表面涂抹标本)AIV核酸,对4份污水H5N1亚型AIV核酸阳性标本进行HA基因RT-PCR扩增和TA克隆测序,测序结果利用Lasergene和Mega 5.0软件进行氨基酸比对和进化树构建.结果AIVH5N1抗体监测结果显示,家禽市场职业暴露人群血清H5N1抗体阳性率为25.5％(26/102),其中乡镇和城区家禽市场职业暴露人群阳性率分别为50.0％(9/18)和25.4％(17/67),乡镇家禽市场职业暴露人群阳性率高于城区.长沙市家禽市场环境中H5亚型AIV核酸阳性率为31.3％(50/160),其中乡镇家禽市场阳性率为37.3％(31/83),高于城区家禽市场24.7％(19/77)；不同标本H5亚型AIV核酸阳性率不同:污水(50.0％,24/48)、羽毛(44.5％,4/9)、禽类粪便(29.8％,14/47)和禽类笼具表面涂抹(14.3％,8/56)；差异均有统计学意义(P＜0.01).TA克隆测序得到4个AIVH5N1亚型HA基因序列,进化树显示4个AIVH5N1亚型HA基因与中国内地和香港禽来源的AIV分离株为同一分组,属于欧亚分支；4个AIV H5N1亚型HA基因受体结合位点氨基酸序列仍然为禽源(QSG)、HA1和HA2蛋白之间连接肽为多个碱性氨基酸序列(RERRRKK或RERRGKK),与入源AIV H5N1亚型具有相同的受体结合位点和高致病性的分子特征.结论 长沙市家禽市场环境中存在较多数量的AIVH5N1亚型,是导致职业暴露人群抗体阳性率达25.5％的原因之一；环境中存在的AIVH5N1亚型HA基因表现出来的高致病性分子特征,增加了家禽市场环境中发生AIV传播的风险.
目的 對長沙市傢禽市場職業暴露人群進行禽流感病毒(AIV)H5N1亞型抗體水平和環境AIV覈痠檢測,併對環境中AIV H5N1亞型的血凝素(HA)基因進行測序分析.方法 抽取長沙市1箇區和1箇縣,各選擇2箇城區或鄉鎮傢禽市場進行職業暴露人群H5N1抗體和環境AIV覈痠檢測.利用單放射免疫擴散溶血實驗(SRH)對102份傢禽市場職業暴露人員血清標本進行H5N1抗體檢測,real-time PCR方法檢測160份傢禽市場環境標本(汙水、禽類糞便、羽毛和禽類籠具錶麵塗抹標本)AIV覈痠,對4份汙水H5N1亞型AIV覈痠暘性標本進行HA基因RT-PCR擴增和TA剋隆測序,測序結果利用Lasergene和Mega 5.0軟件進行氨基痠比對和進化樹構建.結果AIVH5N1抗體鑑測結果顯示,傢禽市場職業暴露人群血清H5N1抗體暘性率為25.5％(26/102),其中鄉鎮和城區傢禽市場職業暴露人群暘性率分彆為50.0％(9/18)和25.4％(17/67),鄉鎮傢禽市場職業暴露人群暘性率高于城區.長沙市傢禽市場環境中H5亞型AIV覈痠暘性率為31.3％(50/160),其中鄉鎮傢禽市場暘性率為37.3％(31/83),高于城區傢禽市場24.7％(19/77)；不同標本H5亞型AIV覈痠暘性率不同:汙水(50.0％,24/48)、羽毛(44.5％,4/9)、禽類糞便(29.8％,14/47)和禽類籠具錶麵塗抹(14.3％,8/56)；差異均有統計學意義(P＜0.01).TA剋隆測序得到4箇AIVH5N1亞型HA基因序列,進化樹顯示4箇AIVH5N1亞型HA基因與中國內地和香港禽來源的AIV分離株為同一分組,屬于歐亞分支；4箇AIV H5N1亞型HA基因受體結閤位點氨基痠序列仍然為禽源(QSG)、HA1和HA2蛋白之間連接肽為多箇堿性氨基痠序列(RERRRKK或RERRGKK),與入源AIV H5N1亞型具有相同的受體結閤位點和高緻病性的分子特徵.結論 長沙市傢禽市場環境中存在較多數量的AIVH5N1亞型,是導緻職業暴露人群抗體暘性率達25.5％的原因之一；環境中存在的AIVH5N1亞型HA基因錶現齣來的高緻病性分子特徵,增加瞭傢禽市場環境中髮生AIV傳播的風險.
목적 대장사시가금시장직업폭로인군진행금류감병독(AIV)H5N1아형항체수평화배경AIV핵산검측,병대배경중AIV H5N1아형적혈응소(HA)기인진행측서분석.방법 추취장사시1개구화1개현,각선택2개성구혹향진가금시장진행직업폭로인군H5N1항체화배경AIV핵산검측.이용단방사면역확산용혈실험(SRH)대102빈가금시장직업폭로인원혈청표본진행H5N1항체검측,real-time PCR방법검측160빈가금시장배경표본(오수、금류분편、우모화금류롱구표면도말표본)AIV핵산,대4빈오수H5N1아형AIV핵산양성표본진행HA기인RT-PCR확증화TA극륭측서,측서결과이용Lasergene화Mega 5.0연건진행안기산비대화진화수구건.결과AIVH5N1항체감측결과현시,가금시장직업폭로인군혈청H5N1항체양성솔위25.5％(26/102),기중향진화성구가금시장직업폭로인군양성솔분별위50.0％(9/18)화25.4％(17/67),향진가금시장직업폭로인군양성솔고우성구.장사시가금시장배경중H5아형AIV핵산양성솔위31.3％(50/160),기중향진가금시장양성솔위37.3％(31/83),고우성구가금시장24.7％(19/77)；불동표본H5아형AIV핵산양성솔불동:오수(50.0％,24/48)、우모(44.5％,4/9)、금류분편(29.8％,14/47)화금류롱구표면도말(14.3％,8/56)；차이균유통계학의의(P＜0.01).TA극륭측서득도4개AIVH5N1아형HA기인서렬,진화수현시4개AIVH5N1아형HA기인여중국내지화향항금래원적AIV분리주위동일분조,속우구아분지；4개AIV H5N1아형HA기인수체결합위점안기산서렬잉연위금원(QSG)、HA1화HA2단백지간련접태위다개감성안기산서렬(RERRRKK혹RERRGKK),여입원AIV H5N1아형구유상동적수체결합위점화고치병성적분자특정.결론 장사시가금시장배경중존재교다수량적AIVH5N1아형,시도치직업폭로인군항체양성솔체25.5％적원인지일；배경중존재적AIVH5N1아형HA기인표현출래적고치병성분자특정,증가료가금시장배경중발생AIV전파적풍험.
Objective To investigate the risk of H5Nl subtype avian influenza virus (AIV)transmission in the poultry market environment in Changsha city.H5N1 antibody levels among the groups related occupational exposure and AIV nucleic acid in the environment of poultry markets were detected.The characteristics of hamagglutinin (HA) genes of H5N1 AIV in the environment were analyzed.Methods One district and one county from Changsha city were selected randomly and two poultry markets at inner city or township levels were selected in the same district or county respectively.H5N1 antibody of the occupational exposure groups in the poultry market was tested and AIV nucleic acid in the poultry market environnent monitored.One hundred and two blood samples of the occupational exposure groups were tested for H5N1 antibody with single radioimmunoassay diffusion hemolysis (SRH) while 160 environment samples(from sewage,birds stools,feathers and smearing samples of poultry cages) in the poultry market were also detected for AIV nucleic acid with real-time PCR method.Four sewage samples of H5N1 subtype AIV were collected from poultry markets in Changsha,and the HA genes of H5N1 subtype AIV amplified by RT-PCR and then sequenced with TA cloning.Amino acid sequence alignment and phylogenetic tree analysis were conducted by Lasergene and Mega 5.0 software.Results The results through H5N1 antibody monitoring program showed that H5N1 antibody positive rates from workers were 25.5％ (26/102),50.0％ (9/18) and 25.4％ (17/67) respectively in the poultry markets of township and inner cities.H5N1 antibody positive rate in the township poultry markets was higher than in the inner cities poultry markets.Results from the surveillance on AIV nucleic acid showed that the overall H5 subtype positive rate in Changsha poultry markets was 31.3％ (50/160),and the positive rate of townships poultry markets was 37.3％ (31/83 ),which were both higher than those from the inner cities poultry markets (24.7％,19/77).H5 subtype AIV positive rate was different in the tested specimens,with ranking of positive rates were sewage (50.0％,24/48),feathers (44.5％,4/9 ),birds stools ( 29.8％,14/47) and smearing samples of poultry cages (14.3％,8/56),with statistically significant differences (P＜0.01).Four H5N1 HA genes TA cloning were successfully constructed and identified as Eurasian branch,similar to viruses isolated in mainland China and Hong Kong in the same group,according to genetic analysis.Sequence data of the four HA genes showed the same feature of high pathogenicity,compared to the H5N 1 AIV from mainland China of human origin.The receptor specificities were still with avian influenza origin (QSG)and the connecting peptide between HA1 and HA2 possessing the polybasic motif (RERRRKK or RERRGKK).Conclusion One of the reasons for H5N1 antibody positive rate of 25.5％ among poultry markets workers was that there were large numbers of H5N 1 subtype AIV detected in the environment of poultry markets and HA genes of H5N 1 subtype AIV in the poultry markets environment carried molecular characteristics of highly pathogenic which could increase the risk for H5N1 subtype AIV transmission in the environment of poultry markets.
