Laboratory Animal and Comparative Medicine ›› 2017, Vol. 37 ›› Issue (1): 32-35.DOI: 10.3969/j.issn.1674-5817.2017.01.007

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Development of Dual PCR for Detection of Rat Parvovirus

RAO Dan, ZHU Yu-jun, WU Miao-li, YUAN Wen, WANG Jing, YIN Xue-qin, CONG Feng, LIAN Yue-xiao, HUANG Bi-hong, XU Feng-jiao, LIU Xiang-nan, LIU Zhu-hong, HUANG Ren, ZHANG Yu, GUO Peng-ju   

  • Received:2016-09-05 Online:2017-02-25 Published:2017-02-25

Abstract: Objective To establish a PCR method for rapidly detecting rat parvovirus. Methods According to the characters of nucleotide sequence of rat parvovirus, a dual PCR was developed to differentially detect RPV and other three rat parvovirus H-1/KRV/RMV. A pair of primers amplifying VP2 gene were applied to exclusively amplify RPV while another pair of primers targeting the NS1 gene was designed for specific amplification of H-1, KRV and RMV primer. Result Rat parvovirus were screened without detecting minute virus of mice which had high nucleotide homology with rat parvovirus and also negative for three other microorganism pathogen. Sensitivity tests showed that the minimum detectable concentration was as low as 1 000 copies μL. When dual PCR combined with sequencing were applied to detect 23 clinical samples, seven samples detected were RMV positive including one coinfected with RPV. Conclusion The dual PCR was verified to be specific and sensitive which could be used as a reliable method to screen rat parvovirus.

Key words: Rat Parvovirus (RPV), H-1, KRV, RMV, Dual PCR

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