Abstract：Detection of bacteria with traditional method of microscope examination leads to high inaccuracy .On the contrary , the PCR method is specific and accurate to identify bacterial species . A multiplex PCR method for accurate detection of conventional bacteria in aquatic animals was developed and used to amplify Hly A gene in Aeromonas hydrophila ,Khe gene in Klebsiella pneumoniae ,Sip gene in Streptococcus agalactiae ,Ser C gene ,Eip gene in Edwardsiella ictaluri ,and Muk F gene ,and Gad B gene in Edwardsiella tarda .The results showed that the amplification was found to be high specificity .A double PCR method for simultaneous detection of S . agalactiae and E .ictalur and a multiplex PCR method for simultaneous detection of A . hydrophila , K . pneumoniae , E . tarda and E .ictalur were developed .In order to verify the accuracy of the multiplex PCR method , the diseased ricefield eel (Monopterus albus)were collected for isolation and culture of bacteria ,as well as for DNA extraction from the cultured bacteria .K .pneumoniae and A .hydrophila were detected by multiplex PCR method .And the established multiplex PCR had important significance for rapid detection of aquatic bacterial conventional pathogenic bacterium and molecular epidemiological research .
收稿日期: 2015-05-25
引用本文:
艾克特,徐佩,周静文,万晶,杨代勤,许巧情. 5 种淡水细菌的多重 PC R 检测方法的建立及其应用[J]. 水产科学, 2015, 34(5): 321-326.
AI Kete,XU Pei,ZHOU Jingwen,WAN Jing,YANG Daiqin ,XU Qiaoqing . Establishment and Application of Multiplex PCR Assay for Detection of Five Freshwater Bacterial Species. Fisheries Science, 2015, 34(5): 321-326.
