Baloda S B, Krovacek K, Eriksson L, Linné T, Månsson I
Section of Bacteriology and Epizootology, Faculty of Veterinary Medicine, Swedish University of Agricultural Sciences, Uppsala.
Comp Immunol Microbiol Infect Dis. 1995 Jan;18(1):17-26. doi: 10.1016/0147-9571(94)e0001-a.
A polymerase chain reaction (PCR) technique was used to assay the presence of the aerolysin gene in a total of 89 Aeromonas hydrophila and A. sobria strains isolated from drinking water, fish and foods. These strains were also characterized for the production of virulence factors such as haemolysin, protease and cytotoxin. The primers used in the PCR targeted a 209-bp fragment of the aer gene coding for the beta-haemolysin and detected template DNA only in haemolytic A. hydrophila strains. The cell-free culture supernatants of these aerolysin-positive A. hydrophila strains were also cytotoxic to the HeLa and McCoy cells. The haemolytic A. sobria and non-haemolytic A. hydrophila were consistently negative in the PCR assay. Primer specificity was determined in the PCR by using a control haemolytic Escherichia coli, Streptococcus pyogenes and a restriction endonuclease assay. The PCR clearly identified the aerolysin-producing strains of A. hydrophila and may have application as a rapid species-specific virulence test.
采用聚合酶链反应(PCR)技术检测了从饮用水、鱼类和食品中分离出的89株嗜水气单胞菌和温和气单胞菌中溶血素基因的存在情况。还对这些菌株产生溶血素、蛋白酶和细胞毒素等毒力因子的情况进行了鉴定。PCR中使用的引物靶向编码β-溶血素的aer基因的一个209碱基对片段,并且仅在溶血性嗜水气单胞菌菌株中检测到模板DNA。这些溶血素阳性嗜水气单胞菌菌株的无细胞培养上清液对HeLa细胞和McCoy细胞也具有细胞毒性。溶血性温和气单胞菌和非溶血性嗜水气单胞菌在PCR检测中始终呈阴性。通过使用对照溶血性大肠杆菌、化脓性链球菌和限制性内切酶分析在PCR中确定引物特异性。PCR能清晰地鉴定出产生溶血素的嗜水气单胞菌菌株,可能可用作一种快速的种特异性毒力检测方法。