沙门菌变性高效液相色谱法鉴定和分型研究  

IDENTIFICATION AND TYPING OF SALMONELLA USING PCR-DHPLC METHOD

作　　者：杨福江[1] 王玉平[1] 吴永宁[2] 沈建忠[3] 

机构地区：[1]河北邢台医学高等专科学校,邢台054000 [2]中国疾病预防控制中心营养与食品安全所 [3]中国农业大学动物医学院

出　　处：《现代预防医学》2011年第4期721-724,共4页Modern Preventive Medicine

基　　金：中国农业大学沈建忠教授承担的"十一五"国家科技支撑计划"食品安全关键技术"课题(No.2006BAK02A03)资助

摘　　要：[目的]建立PCR与变性高效液相色谱(DHPLC)相结合的方法,对沙门菌进行鉴定和分型。[方法]针对16S rRNA基因或16S-23S之间序列保守区设计3对引物S1、S2和S3,PCR扩增产物或杂交产物用变性高效液相色谱仪鉴定。[结果]柱温50℃时,引物S1产物在不同血清型沙门菌显示为特异DHPLC图谱;柱温61.4℃时,引物S2杂交产物在不同血清型沙门菌显示为特异DHPLC图谱;引物S3可用于沙门菌的特异鉴定,但不能区分血清型;同一血清型内各菌株基因高度保守。[结论]PCR-DHPLC方法适用于不同血清型沙门菌的分型,但不适于同一血清型内的菌株分型。[Objective] To establish a PCR-DHPLC（denaturing high-performance liquid chromatography） method for the identification and genotype of different Salmonella serovars.[Methods] Three primer sets（S1,S2 and S3） for the conserved region of 16S rRNA gene or 16S-23S ITS were designed and used for the PCR amplification.The PCR products or hybridization products were detected by DHPLC.[Results] The PCR products for primer S1 were shown as specific peak profiles in different Salmonella serovars at an oven temperature of 50 ℃.The hybridization products for primer S2 were shown as specific peak profiles in three Salmonella serovars at an oven temperature of 61.4 ℃.Primer S3 could be used for the identification of Salmonella spp.None of the three primer sets could be used for the molecular genotype of Salmonella strains.[Conclusion] The PCR-DHPLC method could be used for quickly identification of different Salmonella serovars,but not for genotype of Salmonella strains.

关 键 词：变性高效液相色谱 沙门菌 鉴定 分型 16S RRNA基因 

分 类 号：R378.22[医药卫生—病原生物学]