鸭巴泰病毒抗体间接ELISA检测方法的建立及部分地区的初步流行病学调查
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摘要
为建立检测新发现的鸭巴泰病毒(BATV)感染的血清学方法及初步了解相关地域BATV的流行情况,本研究以浓缩纯化的BATV ZJ-01株作为包被抗原,通过优化反应条件,建立了BATV抗体的间接ELISA检测方法。结果表明,该方法仅与BATV阳性血清发生特异性反应,而对禽流感病毒等8种常见鸭病毒阳性血清无交叉反应,表明其特异性强;经敏感性试验测定,阳性血清1∶320倍稀释时检测仍为阳性,显示其敏感性高。批内重复试验变异系数低于5%,批间重复试验变异系数低于10%,证明其重复性良好。经30份血清样品的比对检测结果显示,该方法与中和试验符合率为100%。应用该方法对采自闽浙赣三省发生产蛋下降种(蛋)鸭群、生长不良肉鸭群共493份血清样品分别检测,结果总阳性率高达29.41%。本研究为BATV的流行病学调查提供了一种特异、敏感、快速的血清学检测方法。
To development serological method for detection of antibodies against newly emerged duck Batai virus(BATV), an indirect ELISA were established with purified BATV ZJ-01 strain as coating antigen, and the reaction conditions were optimized.The result show that the established method were specific to detect anti-sera against BATV, but no cross-reaction to the sera against avian influenza virus and other eight kinds of common duck virus. The sensitivity test showed that the positive serum were still positive in 1∶320 times of dilution, which showed high sensitivity of the assay. The variation coefficient of intra-assay was lower than 5% and the inter-assay was lower than 10%, which indicated that the assay was repeatability. Comparison test analysis showed that the compliance rate was 100% between ELISA and neutralization test, based on the titration of 30 serum samples. A total of 493 clinical serum samples from retarded growth of ducks collected in Fujian, Zhejiang and Jiangxi provinces were tested using this method, the positive rate was as high as 29.41% for BATV. The present study provides a specific, sensitive and rapid serological method for the epidemiological investigation of BATV infection in ducks.
To development serological method for detection of antibodies against newly emerged duck Batai virus(BATV), an indirect ELISA were established with purified BATV ZJ-01 strain as coating antigen, and the reaction conditions were optimized.The result show that the established method were specific to detect anti-sera against BATV, but no cross-reaction to the sera against avian influenza virus and other eight kinds of common duck virus. The sensitivity test showed that the positive serum were still positive in 1∶320 times of dilution, which showed high sensitivity of the assay. The variation coefficient of intra-assay was lower than 5% and the inter-assay was lower than 10%, which indicated that the assay was repeatability. Comparison test analysis showed that the compliance rate was 100% between ELISA and neutralization test, based on the titration of 30 serum samples. A total of 493 clinical serum samples from retarded growth of ducks collected in Fujian, Zhejiang and Jiangxi provinces were tested using this method, the positive rate was as high as 29.41% for BATV. The present study provides a specific, sensitive and rapid serological method for the epidemiological investigation of BATV infection in ducks.
引文
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[11]章丽娇,张清水,傅光华,等.一株鸭源巴泰病毒的分离鉴定[A].中国畜牧兽医学会动物传染病学分会第十六次学术研讨会论文集[C].山东济南:中国畜牧兽医学会动物传染病学分会,2015.
[12]Jost H,Bialonski A,Schmetz C,et al.Isolation and phylogenetic analysis of Batai virus,Germany[J].Am J Trop Med Hyg,2011,84(2):241-243.
[13]Hofmann M,Wietholter A,Blaha I,et al.Surveillance of Batai vims in bovines from Germany[J].Clin Vaccine Immunol,2015,22(6):672-673.
[14]章域震,张海林,杨卫红,等.巴泰病毒云南株生物学性状研究[J].中国人兽共患病学报,2013,29(06):527-532.
[15]姬希文,闫丽萍,颜丕熙,等.鸭坦布苏病毒抗体间接ELISA检测方法的建立[J].中国预防兽医学报,2011,33(08):630-634.
[2]王凤田,吕志,王志玉,等.巴泰病毒研究进展[J].中国人兽共患病学报,2009,25(04):365-367.
[3]Briese T,Bird B,Kapoor V,et al.Batai and Ngari viruses:M segment reassortment and association with severe febrile disease outbreaks in East Africa[J].J Virol,2006,80(11):5627-5630.
[4]Hubalek Z.Mosquito-bome viruses in Europe[J].Parasitol Res,2008,103(Suppl 1):S29-S43.
[5]陶三菊,张海林,杨冬荣,等.云南省澜沧江下游地区虫媒病毒的调査研究[J].中华实验和临床病毒学杂志,2003,17(04):24-28.
[6]黄文丽,张海林,侯宗柳,等.云南省西双版纳发热病人血清虫媒病毒抗体调査[J].地方病通报.2001,16(02):29-30.
[7]Nashed N W,Olson J G,El-Tigani A.Isolation of Batai virus(Bunyaviridae:Bunyavirus)from the blood of suspected malaria patients in Sudan[J].Am J Trop Med Hyg,1993,48(5):676-681.
[8]Shcherbakova S A,Golovinskaia O N,Kliueva E V,et al.Anti-arbovirus antibodies positive serum screening in population of Saratov[J],Vopr Virusol,2002,47(3):32-34.
[9]L'Vov S D,Gromashevskii V L,Kanev E F,et al.The circulation of viruses of the California encephalitis and Bunyamwera groups(Bunyaviridae,Bunyavirus)on the northeastern Russian plain[J].Vopr Virusol,1991,36(1):31-34.
[10]Korobeinikova A S,Nafeev A A,Skvortsova T M.Specific markers for the detection of circulation of Tahyna,Inko and Batai viruses(Bunyaviridae,Bunyavirus)in humans,mosquitoes,ticks and cattle of the Ul'ianovsk region[J].Vopr Virusol,2003,48(1):45-46.
[11]章丽娇,张清水,傅光华,等.一株鸭源巴泰病毒的分离鉴定[A].中国畜牧兽医学会动物传染病学分会第十六次学术研讨会论文集[C].山东济南:中国畜牧兽医学会动物传染病学分会,2015.
[12]Jost H,Bialonski A,Schmetz C,et al.Isolation and phylogenetic analysis of Batai virus,Germany[J].Am J Trop Med Hyg,2011,84(2):241-243.
[13]Hofmann M,Wietholter A,Blaha I,et al.Surveillance of Batai vims in bovines from Germany[J].Clin Vaccine Immunol,2015,22(6):672-673.
[14]章域震,张海林,杨卫红,等.巴泰病毒云南株生物学性状研究[J].中国人兽共患病学报,2013,29(06):527-532.
[15]姬希文,闫丽萍,颜丕熙,等.鸭坦布苏病毒抗体间接ELISA检测方法的建立[J].中国预防兽医学报,2011,33(08):630-634.