Characterization of Functional furVs as a Biomarker for Detection of Aquatic Pathogen Vibrio splendidus

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• 作者：Liang Weikang ; Zhang Shanshan ; Zhang Weiwei ; Li Chenghua and Han Qingxi
• 刊名：Journal of the World Aquaculture Society
• 出版年：2016
• 出版时间：August 2016
• 年：2016
• 卷：47
• 期：4
• 页码：577-586
• 全文大小：674K
• ISSN：1749-7345

文摘

Vibrio splendidus is an important opportunistic pathogen that can infect a broad range of aquatic animals including Apostichopus japonicus, leading to skin ulceration syndrome. In this study, fur gene of V. splendidus (fur_Vs), the gene encoding ferric uptake regulator (Fur) of V. splendidus, was cloned and characterized. fur_Vs possessed the signature ammonia acids of Vibrio sp., and its mRNA level was regulated by iron and coelomic fluid of A. japonicus. While the amino acid sequence of the fur_Vs showed high homology with the compared Fur proteins of other Vibrio sp., the nucleotide sequence of fur_Vs differed greatly from that of the other fur genes from Vibrio sp., based on which a pair of specific primers VSFurRTF3 and VSFurRTR3, was designed. A distinct DNA band with a length of 223 bp can be amplified from V. splendidus with great specificity. The detection limits for V. splendidus in seawater, homogenized tissues of epidermis, muscles, and coelomic fluid from A. japonicus, was 1 × 10², 1 × 10⁵, 1 × 10³, and 1 × 10² CFU/mL, respectively. Moreover, the detection limit was as low as 8 × 10⁻³ pg/μL when using DNA of V. splendidus as a template. Our results provide a simple and rapid polymerase chain reaction method for the sensitive and specific detection of V. splendidus.