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鸡白痢沙门菌等位基因特异性PCR检测方法的建立
引用本文:耿士忠,潘志明,蒋春红,张辉,李求春,焦新安.鸡白痢沙门菌等位基因特异性PCR检测方法的建立[J].中国兽医科技,2007,37(2):113-116.
作者姓名:耿士忠  潘志明  蒋春红  张辉  李求春  焦新安
作者单位:扬州大学江苏省人兽共患病学重点实验室,江苏扬州225009
基金项目:国家重点基础研究发展计划(973)项目(2006CB504400);国家自然科学基金资助项目(30425031);教育部FANEDD专项资金项目(200358)
摘    要:根据鸡白痢沙门菌与鸡伤寒沙门菌的rfbS基因在第237和第598位碱基的不同,设计和合成了等位基因特异性PCR引物,建立了快速检测鸡白痢沙门菌的PCR方法,并应用该方法对鸡白痢沙门菌临床分离样品进行了PCR鉴定。结果显示,该PCR方法能够特异性地鉴定鸡白痢沙门菌,检测灵敏度达100PgDNA。对35个经常规方法鉴定的鸡白痢沙门菌分离株应用等位基因特异性PCR方法进行鉴定,鉴定出33株鸡白痢沙门菌,符合率为94.3%。表明,建立的等位基因特异性PCR方法能够准确而快速地鉴定鸡白痢沙门菌。

关 键 词:鸡白痢沙门菌  等位基因特异性PCR  rfbS基因
文章编号:1673-4696(2007)02-0113-04
收稿时间:2006-09-21
修稿时间:2006-12-12

Development of allele-specific PCR assay for detecting Salmonella pullorum
GENG Shi-zhong, PAN Zhi-ming,JIANG Chun-hong, ZHANG H ui, LI Qiu-chun,JIAO Xin-an.Development of allele-specific PCR assay for detecting Salmonella pullorum[J].Chinese Journal of Veterinary Science and Technology,2007,37(2):113-116.
Authors:GENG Shi-zhong  PAN Zhi-ming  JIANG Chun-hong  ZHANG H ui  LI Qiu-chun  JIAO Xin-an
Abstract:Based on the nuclei polymorphism of rfbS gene sequence of Salmonella pullorum at site 237 and 598 compared with S. gallinarum, a pair of allele-specific primers were designed and synthesized, and an allele-specific PCR method for detecting S. pullorum was developed. The PCR detection result of 35 S. pullorum clinical isolates was 94.3 %0 consistent with result of the serological and biochemical method and the sensitivity of the PCR assay was 100 pg. The results showed that the developed allele-specific PCR is a very rapid, sensitive and specific molecular tool for the detection and identification of S. pullorum isolates.
Keywords:Salmonella pullorum  allele-specific PCR  rfbS gene
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