| 牛传染性鼻气管炎病毒PCR检测方法的建立及初步应用 |
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| 引用本文： | 王嵩,林红丽,王宇鹏,等.牛传染性鼻气管炎病毒PCR检测方法的建立及初步应用[J].黑龙江八一农垦大学学报,2014(3):26-29. |
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| 作者姓名： | 王嵩 林红丽 王宇鹏 等 |
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| 作者单位： | [1]黑龙江八一农垦大学动物科技学院,大庆163319; [2]黑龙江八一农垦大学生命科学技术学院,大庆163319; |
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| 基金项目： | 黑龙江省教育厅项目（12521372）;国家科技支撑计划课题（2012BAD12B05）. |
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| 摘 要： | 为建立一种快速诊断牛传染性鼻气管炎病毒(IBRV)病的方法,根据GenBank中登录的IBRV保守基因gB序列,利用分子生物学软件Primer5.0设计1对特异引物,优化反应体系后,建立了IBRV PCR检测方法。特异性试验结果显示该方法可从IBRV DQ株中扩增出398 bp的特异性片段,而对牛呼吸道合胞体病毒、牛副流感病毒、MDBK细胞的扩增结果均为阴性。该方法检测IBRV的敏感性可达10-3 TCID50·mL-1。应用建立的PCR方法能够从发病的临床样品中检测出IBRV且比病毒分离方法更为敏感,操作简便。结果表明建立的PCR方法具有特异、敏感、快速的特点,可用于IBRV的临床检测。
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| 关 键 词： | 牛传染性鼻气管炎病毒 聚合酶链反应 检测 |
Establishment and Application of Single PCR Method of IBRV Infection Diagnosis |
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| Institution: | Wang Song, Lin Hongli, Wang Yupeng, Liu Zhenge, Wang Jie, Yang Mingfa, Yu Liyun, Hou Xilin (1. College of Animal Science and Technology, Heilongjiang Bayi Agricultural University, Daqing 163319; 2. College of Life Science and Technology, Heilongjiang Bayi Agricultural University) |
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| Abstract: | In order to detect IBRV simultaneously,single PCR was established and optimized for IBRV infection. According to the IBRV gB gene sequence in GenBank,one set of primers were designed by Primer5.0.optimization of reaction system,and PCR method for detection of IBRV was established. PCR for specificity test could detect the 369 bp specific fragment from IBRV DQ strain,but it was negative for BRSV,BPIV-3 and MDBK cells. The sensitivity of PCR assay was 10-3 TCID50＆#183;mL-1. PCR method could detect the IBRV from the onset of clinical samples and was more sensitive and easy operation than virus isolation method. PCR assay was the practical and reliable tool for the diagnosis of IBRV infection. |
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| Keywords: | IBRV single PCR detection |
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