| 牛精液中基因I型牛病毒性腹泻病毒荧光探针定量RT-PCR检测方法的建立 |
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| 引用本文： | 季新成,黄玲,段晓东,员丽娟,牛国辉,于学辉,曾新强,张彦明.牛精液中基因I型牛病毒性腹泻病毒荧光探针定量RT-PCR检测方法的建立[J].中国预防兽医学报,2011,33(1). |
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| 作者姓名： | 季新成 黄玲 段晓东 员丽娟 牛国辉 于学辉 曾新强 张彦明 |
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| 作者单位： | 1. 新疆出入境检验检疫局,新疆,乌鲁木齐,830063
2. 西北农林科技大学动物医学院,陕西,杨凌,712100 |
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| 基金项目： | 国家质量监督检验检疫总局项目基金 |
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| 摘 要： | 为建立牛精液中基因I型牛病毒性腹泻病毒(BVDV-1)的快速检测方法,本研究采用Sephycral S-400凝胶对牛精液过滤处理后提取病毒核酸,根据BVDV-1 5'UTR保守区基因序列,设计特异性引物和荧光探针,通过反应条件的优化,建立了牛精液中BVDV-1荧光定量RT-PCR检测方法.该方法可以检测到牛精液中含量为0.0125 TCID50的病毒,灵敏度比病毒分离方法高200倍～2000倍,比常规RT-PCR方法高10倍.对从同一个牛场6个月内采集的120份新鲜牛精液和40份冷冻牛精液用该荧光RT-PCR方法检测,没有检测到BVDV-1阳性样品.对其中的10头牛定期检测精液中病毒的同时,并同步检测了全血中的病毒和血清抗体,结果血清抗体阳性牛精液中没有检测到病毒,本研究结果表明,不能以血清抗体阴阳性做为精液是否带毒的依据.
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| 关 键 词： | 牛精液 基因I型BVDV 荧光定量RT-PCR检测 血清抗体检测 |
Establishment of fluorescent quantitative RT-PCR for genotype I BVDV detection in bovine semen |
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| JI Xin-cheng,HUANG Ling,DUAN Xiao-dong,YUAN Li-juan,NIU Guo-hui,YU Xue-hui,ZENG Xin-qiang,ZHANG Yan-ming.Establishment of fluorescent quantitative RT-PCR for genotype I BVDV detection in bovine semen[J].Chinese Journal of Preventive Veterinary Medicine,2011,33(1). |
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| Authors: | JI Xin-cheng HUANG Ling DUAN Xiao-dong YUAN Li-juan NIU Guo-hui YU Xue-hui ZENG Xin-qiang ZHANG Yan-ming |
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| Institution: | JI Xin-cheng1,HUANG Ling1,DUAN Xiao-dong1,YUAN Li-juan1,NIU Guo-hui1,YU Xue-hui1,ZENG Xin-qiang1,ZHANG Yan-ming2*(1.Xinjiang Exit-import Inspection and Quarantine Bureau,Urumqi 830063,China,2.College of Veterinary Medicine,Northwest A&F university,Yangling 712100,China) |
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| Abstract: | A fluorescent quantitative RT-PCR(FQ-RT-PCR) method was established for direct detection of genotype I BVDV in bovine semen using specific primers and probe derived from the BVDV 5' UTR region.BVDV RNA was extracted from raw bovine semen or extended bovine semen samples by including a sephacryl S-400 chromatography filtration step to remove inhibitors.The FQ-RT-PCR assay could detect 0.0125 TCID50 BVDV which was 200 to 2,000 times more sensitive than virus isolation method,and 10-fold more sensitive than co... |
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| Keywords: | Bovine semen genotype I BVDV fluorescent quantitative RT-PCR detection serum antibodies detection |
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