Salmonella is a zoonotic important foodborne pathogen，a sensitive dual-PCR detection method was established is beneficial to achieve the detection of pathogenic Salmonella rapidly． In this thesis， the pathogenicity island genes of pathogenic Salmonella were as the research object and two pairs of primers which named Salmonella pathogenicity island mgtC and sopB were designed and synthesized based on the sequence of the Salmonella pathogenicity island genes． The nucleic acid of Salmonella typhimurium ( ATCC9150) was screened as a template，after primer specificity test， sensitivity test of DNA template and optimize the reaction conditions， the double PCR method of rapid differential detection for pathogenic Salmonella was successfully established． Specific test results showed that the primers mgtC and sopB of double PCR only can amplify two specific fragments of the Salmonella typhimurium ( ATCC9150) whose size is 500 bp and 1 000 bp， respectively． The sensitivity results showed that the limit of detection was 10 cfu /mL ． This double PCR method established could identify the virulence factor in Salmonella，which can achieve the detection of pathogenic Salmonella rapidly in high specificity and sensitivity．
Ouyang Ben
,
Sun Zhen
,
Qi Kezong
,
Wang Xueyan
,
Xue Xiuheng
,
Tu Jian
. Double PCR Method for Detection Genes mgtC and sopB of Salmonella typhimurium Pathogenicity Island[J]. Acta Agriculturae Boreali-Sinica, 2013
, 28(3)
: 48
-52
.
DOI: 10.3969/j.issn.1000-7091.2013.03.009
[1] 彭海滨． 我国沙门氏菌污染分布概况[J]． 中国国境卫生检疫杂志， 2006， 29( 2) : 125 - 128．
[2] 邹兰，祁克宗，薛秀恒，等． 病性大肠埃希菌和沙门菌毒力岛基因的双重PCR 检测方法的建立[J]． 中国微生态学杂志， 2012， 24( 6) : 504 - 507．
[3] 杨小鹃，吴清平，张菊梅，等． 畜禽肉沙门氏菌和大肠杆菌O157 多重PCR 检测研究[J]． 微生物学通报，2008， 35( 3) : 470 - 474．
[4] Chiu T H，Chen T R． Sequencing of an internal transcribed spacer region of 16S-23S rRNA gene and designing of PCR primers for the detection of Salmonella spp．in food[J]． International Journal of Food Microbiology，2005， 97( 3) : 259 - 265．
[5] Kong R Y C，Lee S K Y，Law T W F， et al． Rapid detected of six types of bacterial pathogens in marine waters by multiplex PCR[J]． Water Research，2002，36 (11):280-2812．
[6] Bustamante V H，Martinez L C，Santana F J， et al． HilDmediated transcriptional cross-talk between SPI-1 andSPI-2[J]． Proceedings of the National Academy of Sciences of the United States of America，2008，105 ( 38) :14591 - 14596．
[7] Matsui M，Takaya A，Yamamoto T． Sigma32-mediated negative regulation of Salmonella pathogenicity island 1 expression[J]． Journal of Bacteriology， 2008， 190( 20) :6636 - 6645．
[8] Suarez M，Russmann H． Molecular mechanisms of Salmonella invasion: the typeⅢ secretion system of the pathogenicity island 1[J]． International Microbiology，1998，1( 3) : 197 - 204．
[9] Daniel B L，Silvane M M，Mario S， et al． Trypanosoma cruzi:Multiplex PCR to detect and classify strains according to groups I and Ⅱ[J]． Experimental Parasitology， 2009，123( 4) : 283 - 291．
[10] Bonnet C，Diarra MS， et al． Pathotype and Antibiotic Resistance Gene Distributions of Escherichia coli Isolates from Broiler Chickens Raised on Antimicrobial-Supplemented Diets [J]． Applied and Environmental Microbiology，2009， 75( 22) : 6955 - 6962．
[11] Juhas M，Crook D W． Genomic islands: tools of bacterial horizontal gene transfer and evolution[J]． Fems Microbiology Reviews， 2009， 33( 2) : 376 - 393．
[12] Marcus S L，Brumell J H，Pfeifer C G． et al． Salmonella pathogenicity islands: big virulence in small packages[J]． Microbes and Infection， 2000，2(2):145-156．
[13] 田菊霞，解宇，杨艳宏． 使用免疫BMPs 纯化细菌PCR反应模板的比较[J]． 科技通报， 2012， 28( 5) : 80 -83．
[14] 王慧，朱瑞良，谭燕玲． 多重PCR 检测三种重要食源性致病菌方法的建立及应用[J]． 中国农业科学，2011， 44( 11) : 2334 - 2340．
[15] Kevin J，Samul Y，Helen Won， et al． Application of a 16S rRNA PCR-High-Resolution Melt Analysis Assay for Rapid Detection of Salmonella Bacteremia[J]． Journal of clinical microbiology， 2011， 50( 3) : 1122 - 1124．
[16] 冯飞，谢振文，曾慕衡． 鼠伤寒沙门氏菌多重PCR检测方法的研究[J]． 中国生物工程杂志，2011，31( 1) : 65 - 69．
[17] 范宏英，吴清平，吴若菁，等． 饮用水中5 种致病菌多重PCR 技术检测研究[J]． 微生物学通报，2005，32:102 - 107．
