[1]陈 艳,辛晓光,杨焕良,等.猪流感抗体间接ELISA检测方法的建立[J].中国预防兽医学报,2007,(04):308-311.
 CHEN Yan,XIN Xiao-guang,YANG Huan-liang,et al.Establishment of an indirect ELISA for detection of antibodies against swine influenza virus[J].Chinese journal of preventive veterinary medicine,2007,(04):308-311.
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猪流感抗体间接ELISA检测方法的建立
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2007年04期
页码:
308-311
栏目:
诊断和检测技术
出版日期:
2007-04-01

文章信息/Info

Title:
Establishment of an indirect ELISA for detection of antibodies against swine influenza virus
文章编号:
1008-0589(2007)04-0308-04
作者:
陈 艳辛晓光杨焕良李 婷李海燕乔传玲陈化兰*
中国农业科学院哈尔滨兽医研究所 农业部动物流感重点开放实验室/兽医生物技术国家重点实验室,黑龙江 哈尔滨 150001
Author(s):
CHEN Yan XIN Xiao-guang YANG Huan-liang LI Ting LI Hai-yan QIAO Chuan-ling CHEN Hua-lan*
Animal Influenza Laboratory of Ministry of Agriculture, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, China
关键词:
猪流感猪流感病毒间接ELISA
Keywords:
swine influenza swine influenza virus indirect ELISA
分类号:
S852.659.5;S854.44
文献标志码:
A
摘要:
猪流感病毒A/Swine/Fujian/668/2001(H3N2)株感染的鸡胚尿囊液,经差速离心后,再经蔗糖密度梯度离心,提纯、纯化的猪流感病毒经NP-40处理并反复冻融,作为猪流感间接ELISA抗原,确立了间接ELISA检测方法。对29份HI试验猪流感为阴性的血清进行了检测,经统计学分析,确定间接ELISA判定标准,被检血清OD490nm值≥0.20判定为阳性。该方法对猪瘟等11种猪疫病阳性血清无交叉反应,批内和批间重复试验的吸收变异系数分别在3.34 % ~8.12 %和6.2 %~9.04 %之间。与HI的符合率达到92.8 %,经卡方检验(P<0.01)比HI试验敏感。为猪流感抗体检测提供了快速、准确、简便的方法。
Abstract:
An indirect ELISA for detecting antibodies against swine influenza virus (SIV) was developed using antigen that was derived from allantoic fluid of chicken embryonated eggs infected with A/Swine/Fujian/668/2001(H3N2). The antigen was purified by differential centrifugation and sucrose density gradient centrifugation, followed by NP-40 treatment and 3 freeze-thaw cycles. Samples with an optical density value≥0.2 were scored positive based on the ELISA data from twenty-nine sera that are known SIV negative in hemagglutination inhibition (HI) test. There was no cross-reactivity detected against virus strains of other 11 swine infectious diseases including classical swine fever. The variant coefficient of optical density value in a batch and between batches varied from 3.34 % to 8.12 %, and from 6.2 % to 9.04 %, respectively. The coincidence rate was 92.8 % between indirect ELISA and HI test. The results showed that indirect ELISA is more sensitive than HI (P<0.01) and could be used as an efficient and sensitive method to detect antibodies against SIV.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期:2006-04-26
作者简介:陈 艳(1976~),女,江苏泰兴人,硕士,主要从事猪流感检测方面的研究.
*通讯作者:E-mail:hlchen@hvri.ac.cn
更新日期/Last Update: 2011-01-30