期刊
JOURNAL OF VETERINARY SCIENCE
卷 11, 期 3, 页码 235-241出版社
KOREAN SOC VETERINARY SCIENCE
DOI: 10.4142/jvs.2010.11.3.235
关键词
Aeromonas salmonicida subsp salmonicida; Flavobacterium psychrophilum; multiplex PCR; Yersinia ruckeri
资金
- Scientific and Technological Research Council of Turkey (TUBITAK) [1070208]
Bacterial cold water disease, enteric red mouth disease and frunculosis are the common bacterial diseases of fish worldwide. The etiologic agents of these diseases are Flavobacterium (F.) psychrophilum, Yersinia (Y.) ruckeri and Aeromonas (A.) salmonicida subsp. salmonicida, respectively. In this study, a multiplex polymerase chain reaction (m-PCR) method with YER8/10-Fer3/4-FP1/3 primer pairs which can identify these fish pathogens simultaneously was developed and optimized. In optimized conditions, neither false specific nor nonspecific amplification occurred. The detection limits of the m-PCR method using DNA extracts from dilutions of pure cultures of bacteria were 35 pg for Y. rucked and F. psychrophilum and 70 pg for A. salmonicida subsp. salmonicida. It was determined that 15 CFU Y. rucked and F. psychrophilum and 30 CFU A. salmonicida subsp. salmonicida could he detected by m-PCR developed using genomic DNA extracted from dilutions of the suspensions. The detection limits in the presence of tissue debris were 125 CFU for Y. ruckeri and F. psychrophilum and 250 CFU for A. salmonicida subsp. salmonicida. In conclusion, we submit that the m-PCR method developed and optimized in this study can be used for accurate and rapid identification of these bacteria.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据