3种淡水鱼病原菌多重PCR检测方法的建立与应用

doi:10.16378/j.cnki.1003-1111.20244

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Abstract A multiplex PCR detection system was developed for three major freshwater pathogens, Aeromonas hydrophila, Edwardsiella tarda and Bacillus cereus, from Liaoning province. hly A, ese D and nhe A were selected for the multiplex PCR. The reaction condition was optimized and the specificity and sensitivity were tested for this method. The results showed that all target fragments were amplified clearly when the concentration of all primer pairs was 0.4 μmol/L, and the concentrations of genomic DNA of A. hydrophila, E. tarda and B. cereus were 2.5, 2.5 and 2.5 ng/μL, as well as dNTP and polymerase were added 2.5 μL and 0.3 μL, respectively, with the annealing temperature of 55 ℃. The sensitivity test showed that the multiplex PCR had a high sensitivity with the detection limit of 4.8 ng for A. hydrophila, 3.0 ng for E. tarda and 2.6 ng for B. cereus. The coincidence rate of the multiplex PCR method and 16S r DNA identification method for suspicious clinical samples was 100%. The established multiplex PCR method was specific, sensitive and rapid to simultaneously detect the three pathogens from freshwater fishes.

Key words： Aeromonas hydrophila Edwardsiella tarda Bacillus cereus multiplex PCR

Received: 10 November 2020

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	HU Zongyun
	YANG Peimin
	YAN Youli

Cite this article:

HU Zongyun,YANG Peimin,YAN Youli. Establishment and Application of Multiplex PCR Assay for Detection of Three Major Freshwater Pathogens[J]. Fisheries Science, 2022, 41(4): 605-613.

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https://www.shchkx.com/EN/10.16378/j.cnki.1003-1111.20244 OR https://www.shchkx.com/EN/Y2022/V41/I4/605

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