| BRSV和BPIV-3双重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,刘振格,王杰,杨明发,余丽芸,侯喜林.BRSV和BPIV-3双重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,2014,26(5):35-38. |
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| 作者姓名： | 王嵩 林红丽 王宇鹏 刘振格 王杰 杨明发 余丽芸 侯喜林 |
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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 |
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| 基金项目： | 黑龙江省教育厅项目，国家科技支撑计划课题 |
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| 摘 要： | 为建立一种快速诊断牛呼吸道合胞体病毒(BRSV),牛副流感病毒3型(BPIV-3)病毒病的方法,根据Genbank中登录的BRSV核衣壳蛋白N基因序列和BPIV-3的核衣壳蛋白N基因序列,利用分子生物学软件Primer5.0设计2对特异引物,经过条件优化,建立了BRSV和BPIV-3的双重PCR检测方法。采用该方法检测BRSV和BPIV-3参考毒株,特异性良好。对参考毒株进行梯度稀释检测,结果该方法检测BRSV的敏感性可达10 TCID50·100μL-1,BPIV-3可达10 TCID50·100μL-1。应用建立的双重PCR方法能够从临床样品中检测出BRSV和BPIV-3。表明建立的双重PCR方法具有特异、敏感、快速的特点,可用于BRSV和BPIV-3的临床检测。
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| 关 键 词： | 牛呼吸道合胞体病毒 牛副流感病毒型 双重反转录聚合酶链反应 |
Establishment and Application of Duplex PCR Assay for BRSV and BPIV-3 Infection |
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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 BRSV and BPIV-3 simultaneously,duplex PCR was established and optimized for BRSV and BPIV-3infection. According to the BRSV N gene sequence and BPIV-3 N gene sequence in GenBank,two sets of primers were designed by Primer5.0,which was used to detect the BRSV and BPIV-3 reference strains and had the good specificity. The sensitivity of duplex PCR assay was 10 TCID50·100 μL^-1for BRSV and BPIV-3. Duplex PCR assay could detect BRSV and BPIV-3 from the clinical samples. The results indicated that duplex PCR was the sensitive and specific method to infect the clinical diagnosis of BRSV and BPIV-3. |
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| Keywords: | BRSV BPIV-3 duplex PCR |
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