利用聚合酶链反应（PCR）技术和平板接种法对美人鱼发光杆菌杀鱼亚种进行简单快速检测。

Simple and rapid detection of Photobacterium damselae ssp. piscicida by a PCR technique and plating method.

作者信息

Rajan P R, Lin J H-Y, Ho M-S, Yang H-L

机构信息

Institute of BioAgricultural Sciences, Academia Sinica, Nankang, Taipei, Taiwan.

出版信息

J Appl Microbiol. 2003;95(6):1375-80. doi: 10.1046/j.1365-2672.2003.02119.x.

DOI:10.1046/j.1365-2672.2003.02119.x

PMID:14633013

Abstract

AIMS

To detect Photobacterium damselae ssp. piscicida using the PCR technique and plating method.

METHODS AND RESULTS

Two strains of P. damselae ssp. piscicida were isolated from cultured cobia (Rachycentron canadum) at two different fish farms in Taiwan. A pair of primers was designed to detect the capsular polysaccharide gene of P. damselae ssp. piscicida by PCR. Reference strains of different genus and different clinical strains were used for this study. The expected product (410 bp) was obtained from both P. damselae ssp. piscicida and P. damselae ssp. damselae, and they were differentiated by culturing on thiosulphate citrate bile salts-sucrose agar (TCBS-1). Photobacterium damselae ssp. damselae grew on TCBS-1 producing green colonies whereas P. damselae ssp. piscicida did not grow.

CONCLUSIONS

The methods used are cost and labour effective when compared with the other methods and commercially available kits.

SIGNIFICANCE AND IMPACT OF THE STUDY

This work provides an integrated set of methods to identify the species P. damselae and to differentiate P. damselae ssp. piscicida from P. damselae ssp. damselae.

摘要

目的

运用聚合酶链反应（PCR）技术和平板接种法检测美人鱼发光杆菌杀鱼亚种。

方法与结果

从台湾两个不同养鱼场养殖的军曹鱼（Rachycentron canadum）中分离出两株美人鱼发光杆菌杀鱼亚种。设计了一对引物，通过PCR检测美人鱼发光杆菌杀鱼亚种的荚膜多糖基因。本研究使用了不同属的参考菌株和不同的临床菌株。从美人鱼发光杆菌杀鱼亚种和美人鱼发光杆菌美人鱼亚种中均获得了预期产物（410 bp），并通过在硫代硫酸盐柠檬酸盐胆盐蔗糖琼脂（TCBS-1）上培养进行区分。美人鱼发光杆菌美人鱼亚种在TCBS-1上生长，产生绿色菌落，而美人鱼发光杆菌杀鱼亚种不生长。