渔业科学进展Issue(4)：99-104,6.DOI:10.11758/yykxjz.20150414
罗氏沼虾（Macrobrachium rosenbergii）幼体病原肠杆菌 PCR 检测技术的建立与应用
Development and Application of the PCR Detection Method of Pathogenic Enterobacters in the Larvae of the Giant Freshwater Prawn, Macrobrachium rosenbergii
摘要
Abstract
The current study was to develop a PCR-based method to detect Enterobacter cloacae and Enterobacter aerogenes in Macrobrachium rosenbergii. Two pairs of primers targeted sequences located within the omp A gene of E. cloacae and gyr B gene of E. aerogenes were used to detect E. cloacae and E. aerogenes. Samples collected from infected larvae were detected with the developed PCR method. The expected DNA fragments of 385 bp and 201 bp were from E. cloacae and E. aerogenes, respectively, and no PCR products were amplified from other bacterium. The sensitivity test showed that the detection limits of PCR were 103 CFU/ml for E. cloacae and 102 CFU/ml for E. aerogenes. In addition, the detection results of larval samples were consistent with the actual case of the infectious disease. In summary, the PCR diagnostic method was specific and sensitive and is a reliable tool for identification of E. cloacae and E. aerogenes in infected samples with a little time and cost, which would play an important role in quick diagnose, epidemiology investigation and SPF populations construction of the giant freshwater prawn.关键词
罗氏沼虾/阴沟肠杆菌/产气肠杆菌/PCR 检测Key words
Macrobrachium rosenbergii/Enterobacter cloacae/Enterobacter aerogenes/PCR detection分类
农业科技引用本文复制引用
陈雪峰,杨国梁,高强,夏正龙,濮剑威,慎佩晶,黄振远..罗氏沼虾（Macrobrachium rosenbergii）幼体病原肠杆菌 PCR 检测技术的建立与应用[J].渔业科学进展,2015,(4):99-104,6.基金项目
国家科技支撑计划项目(2012BAD26B04)、浙江省重大科技专项(2012C12907)、浙江省淡水养殖重点科技创新团队项目(2010R50026-02)和湖州市重大科技专项(2011ZD2005)共同资助。 （2012BAD26B04）
