口蹄疫病毒O、A和AsiaⅠ型分型基因芯片的构建与评价

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Abstract Foot-and-mouth disease (FMD) is an acute, hot, and highly contagious animal disease caused by Foot-and-mouth disease virus (FMDV). This study is to establish a gene chip that distinguishes FMDV serotypes O, A and AsiaⅠ. Three specific primers were designed according to the VP1 gene sequences of FMDV, and three target fragments of 248, 206 and 239 bp were obtained by RT-PCR. Three kinds of probes were designed and labeled with primer markers. A genotyping microarray encoding O, A and AsiaⅠ was established and its sensitivity, specificity, reproducibility and shelf life were evaluated. The optimum reaction conditions were as follows: hydration temperature was 37 ℃, confluent concentration was 0.25% BH4Na, 25% ethanol, and hybridization temperature was 42 ℃. Sensitivity test showed that FMDV-O, FMDV-A and FMDV-AsiaⅠ were 10-5, 10-7, 10-5 after the band is not visible. The PCR detection limits were: FMDV-O 118 pg/mL, 4.3×106 copies/μL; FMDV-A 18.4 pg/mL, 8.1×104 copies/μL; FMDV-AsiaⅠ 129 pg/mL, 4.9×105 copies/μL. The FMDV-O dilution reaches 10-6; the FMDV-A dilution reaches 10-8; the FMDV-AsiaⅠ dilution reaches 10-6, the sensitivity of the method was 10~100 times higher than that of a conventional PCR. There was no cross-hybrid among the detecting surfaces O, A and A1; the chip was reproducible and the prepared chip could be stored for at least three months. In this study, the experimental conditions were optimized, and a fast and simple FMDV classification method was established to provide a new technique for the classification of FMDV.

Key words： Foot-and-mouth disease virus Microarray Typing Screening Construction

Received: 17 July 2017 Published: 04 February 2018

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	Articles by authors
	LIU Zhi-Peng
	OU Yang-Ta
	HUANG Xiao-Bei
	GU Xiang
	DIAO Yu-Jia
	CAO San-Jie
	WEN Yi-Beng
	WU Rui
	DIAO Qi
	WEN Xin-Tian

Cite this article:

LIU Zhi-Peng,OU Yang-Ta,HUANG Xiao-Bei, et al. Construction of a Gene Chip for Differentiating Serotypes O, A and AsiaⅠ of Foot-and-mouth disease virus[J]. , 2018, 26(2): 330-338.

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http://journal05.magtech.org.cn/Jwk_ny/EN/ OR http://journal05.magtech.org.cn/Jwk_ny/EN/Y2018/V26/I2/330

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