无乳链球菌和海豚链球菌早期预警分子检测

doi:10.16378/j.cnki.1003-1111.19191

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Abstract Two pairs of specific primers designed according to the 16S rRNA partial gene sequence of Streptococcus agalactiae and the Sip gene sequence of S. iniae, respectively, were applied to establish a rapid and sensitive PCR detection system, aiming to detect trivial amounts of S. agalactiae and S. iniae simultaneously. Two distinct DNA fragments of 870 bp and 614 bp covering the 16S rRNA partial gene sequence and the Sip gene sequence were amplified from the genomic DNA of S. agalactiae and S. iniae, respectively. No cross-reactivity was observed between the two Streptococcus species or with other fish/nonfish pathogens, including Edwards ellatarda, Aeromonas veronii, Vibrio vulnificus, A. sobria, V. alginolyticus, and Escherichia coli. The two primer pairs were capable of detecting genomic DNA of S. agalactiae and S. iniae in concentrations as low as 9.84×10^-5 ng/μL and 9.30×10^-5 ng/μL, respectively, and of detecting the bacterial cells of S. agalactiae and S. iniae in amount as low as 2.76×10³ cfu/mL and 2.51×10³ cfu/mL. The duplex PCR designed in this study had strong specificity, high sensitivity, high detection rate, and was used for rapid identification of lower amount of S. agalactiae and S. iniae. The finding provides a valuable tool for early warning detection of the pathogens leading to streptococosis.

Key words： tilapia Streptococcus agalactiae S. iniae early warning molecular detection

Received: 05 September 2019

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	Articles by authors
	CUI Miao
	WU Min
	LIU Ru
	LI Jingjing
	ZHANG Huijie
	XU Delin
	ZHANG Qizhong

Cite this article:

CUI Miao,WU Min,LIU Ru, et al. Molecular Detection and Early Warning of Streptococcus agalactiae and S. iniae[J]. Fisheries Science, 2021, 40(4): 589-595.

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https://www.shchkx.com/EN/10.16378/j.cnki.1003-1111.19191 OR https://www.shchkx.com/EN/Y2021/V40/I4/589

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