马麝病毒性出血症RT-PCR诊断方法的建立及初步应用

doi:10.3969/j.issn.1674-7968.2020.05.019

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Abstract Moschus chrysogaster (sifanicus) viral hemorrhagic disease (McVHD) is an acute and highly lethal viral infectious disease, and can be a serious threat to the health and safety of musk deer population in China. McVHD is caused by Moschus chrysogaster hemorrhagic disease virus (McHDV) whose genome is highly homologous with Rabbit hemorrhagic disease virus (RHDV) which belongs to Caliciviridae Lagovirus. To develop a rapid and simple diagnostic method for McVHD, a pair of specific primers McVHD-F/R were designed and synthesized based on the comparative analysis of whole genome sequence of McHDV and RHDV in GenBank. Based on the optimization of reaction condition and analysis of specificity and sensitivity, the reverse transcription PCR (RT-PCR) method for McVHD diagnosis was established. The results showed that a specific 448 bp fragment was amplified from McHDV genome using the established RT-PCR method. The RT-PCR method was highly specific and hypersensitive, and had no cross-reaction?with?other 10 pathogenic microorganisms such as Salmonella, Pasteurella multocida, Klebsiella pneumoniae. The RT-PCR was?able?to?detect?as?low?as?10^-5 ng/μL (2.38×; 10³ copies/μL) of?McHDV RNA.?The detection results?of?clinical?samples?revealed?that?the?diagnostic method was able to effectively and quickly detect viral nucleic acid in clinical samples, and the results were completely consistent with the traditional pathogen separation identification. The RT-PCR method established in this study could provide technical support for diagnosis and molecular epidemiological survey of McVHD.

Key words： Moschus chrysogaster (sifanicus) viral hemorrhagic disease (McVHD) RT-PCR Diagnosis

Received: 08 October 2019

ZTFLH:

S852.62

Corresponding Authors: *bsjdy@126.com

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	WANG Qian
	CHEN Wen-Zhe
	DANG Qing-Qing
	SUN Ji-Wen
	BAO Shi-Jun

Cite this article:

WANG Qian,CHEN Wen-Zhe,DANG Qing-Qing, et al. Establishment and Preliminary Application of RT-PCR Diagnosis Method for Moschus chrysogaster (sifanicus) Viral Hemorrhagic Disease[J]. 农业生物技术学报, 2020, 28(5): 945-950.

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http://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2020.05.019 OR http://journal05.magtech.org.cn/Jwk_ny/EN/Y2020/V28/I5/945

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