| 犬源牛犬细小病毒和犬圆环病毒二联PCR检测方法的建立及应用 |
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| 引用本文： | 于永乐,姚延珠,韩先杰,张传美,秦志华,杨瑞梅,张洪亮,刘维全,单虎.犬源牛犬细小病毒和犬圆环病毒二联PCR检测方法的建立及应用[J].畜牧兽医学报,2022,53(3):978-982. |
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| 作者姓名： | 于永乐 姚延珠 韩先杰 张传美 秦志华 杨瑞梅 张洪亮 刘维全 单虎 |
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| 作者单位： | 1. 青岛农业大学动物医学院 山东省预防兽医学重点实验室, 青岛 266109;2. 青岛农业大学植物医学学院, 青岛 266109;3. 中国农业大学生物学院 农业生物技术国家重点实验室, 北京 100193 |
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| 基金项目： | “十三五”国家重点研发计划资助项目(2016YFD0501001;2016YFD0501004)； |
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| 摘 要： | 本研究旨在建立可同时检测犬源牛犬细小病毒(CBoV)和犬圆环病毒(CCV)的二联PCR检测方法,并对两种病毒病当前的流行情况进行监测和调查.分别将已发表的CBoV和CCV基因组序列进行同源性比对,选择高同源区段,应用Primer Primier 5计算机软件设计并合成了2对特异性扩增引物,目的片段大小分别为170 bp...
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| 关 键 词： | 犬源牛犬细小病毒 犬圆环病毒 二联PCR |
| 收稿时间： | 2021-05-25 |
Establishment of Duplex PCR Detective Techniques for Canine Bocavirus and Canine Circovirus |
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| YU Yongle,YAO Yanzhu,HAN Xianjie,ZHANG Chuanmei,QIN Zhihua,YANG Ruimei,ZHANG Hongliang,LIU Weiquan,SHAN Hu.Establishment of Duplex PCR Detective Techniques for Canine Bocavirus and Canine Circovirus[J].Acta Veterinaria et Zootechnica Sinica,2022,53(3):978-982. |
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| Authors: | YU Yongle YAO Yanzhu HAN Xianjie ZHANG Chuanmei QIN Zhihua YANG Ruimei ZHANG Hongliang LIU Weiquan SHAN Hu |
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| Institution: | 1. College of Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China;2. College of Plant Health and Medicine, Qingdao Agricultural University, Qingdao 266109, China;3. State Key Laboratory of Agrobiotechnology, Department of Biochemistry and Molecular Biology, College of Biological Sciences, China Agricultural University, Beijing 100193, China |
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| Abstract: | This study was conducted to develop the duplex PCR assay for detection simultaneously and discrimination of canine bocavirus (CBoV) and canine circovirus (CCV). The published gene sequences of CBoV and CCV in GenBank were analyzed and compared with each other, respectively. Two pairs of specific primers were designed with the Primer Primier 5 software for PCR amplification. The results showed that two specific fragments of the two viruses were simultaneously amplified and the length of amplified fragments were as follows:170 bp for CBoV and 239 bp for CCV. The sensitivity and specificity tests showed that the assay could detect at least 3.0 pg of target DNA, and there was no cross-reaction with those clinically relevant pathogens, ie. canine parvovirus type 2, canine distemper virus and canine parainfluenza virus. The detection results for 30 clinical samples with the duplex PCR showed that 4 samples were positive for CBoV and 3 for CCV. These results indicated that the method could provide technical support for the detection of CBoV and CCV, and epidemiological investigation of canine viral diarrhea. |
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| Keywords: | canine bocavirus canine circovirus duplex PCR |
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