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

doi:10.11843/j.issn.0366-6964.2022.03.031

Acta Veterinaria et Zootechnica Sinica ›› 2022, Vol. 53 ›› Issue (3): 978-982.doi: 10.11843/j.issn.0366-6964.2022.03.031

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Establishment of Duplex PCR Detective Techniques for Canine Bocavirus and Canine Circovirus

YU Yongle¹, YAO Yanzhu², HAN Xianjie¹, ZHANG Chuanmei¹, QIN Zhihua¹, YANG Ruimei¹, ZHANG Hongliang¹, LIU Weiquan^3*, SHAN Hu^1*

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

Received:2021-05-25 Online:2022-03-23 Published:2022-03-31

Abstract

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.

Key words: canine bocavirus, canine circovirus, duplex PCR

CLC Number:

S852.655

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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Establishment of Duplex PCR Detective Techniques for Canine Bocavirus and Canine Circovirus

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