威海地区两种海洋弧菌多重降落PCR体系的建立与应用

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Abstract Objective To develop a multiplex touchdown PCR system for simultaneous detection Vibrio vulnificus and Vibrio alginolyticus and analyze the detection rate of Marine vibrio in weihai foodborne diseases.Methods The fecal specimens of patients with suspected foodborne diseases were separated, cultured and analyzed,two pairs of specific primers were designed based on vvhA gene of Vibrio vulnificus and toxR gene of Vibrio alginolyticus and detected primer specificity with primer-BLAST.The primers were used to establish multiplex touchdown PCR, and detected 93 fecal specimens, and the results were compared with the bacterial culture method.Results 45 cases of Marine vibrio were detected in fecal samples from patients with suspected foodborne diseases, mainly by the Vibrio parahaemolyticus, followed by Vibrio vulnificus and Vibrio alginolyticus.Multiplex touchdown PCR established two genes of pathogenic bacteria (vvhA, toxR) DNA detection limit were 104 CFU/mL, 103 CFU/mL;Compared with the bacterial culture method, the sensitivity and total sensitivity reached 100%. The specificity of vvhA and toxR gene was 96.47% and 95.45% respectively, and the total specificity was 95.95%.Conclusion The system of multiplex touchdown PCR detection we established can be used in the rapid diagnosis and epidemiological investigation of foodborne infection caused by Vibrio vulnificus and Vibrio alginolyticus.

Key words： Vibrio vulnificus vibrioalginolyticus PCR pathogenic marine vibrio

Received: 16 December 2017

PACS:

R378.3

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	XU Xiaojun
	WU Yulong

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XU Xiaojun,WU Yulong. Establishment and application of multiplex touchdown PCR system for detection of two kinds of Marine vibrio in Weihai region[J]. 滨州医学院学报, 2018, 41(2): 87-89.

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http://manu58.magtech.com.cn/Jwk_bzyxy/EN/ OR http://manu58.magtech.com.cn/Jwk_bzyxy/EN/Y2018/V41/I2/87

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