Virulence genotyping of Salmonella spp. with multiplex PCR
Virulence genotyping of Salmonella spp. with multiplex PCR
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DOI：
10.1637/7417.1
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发表时间：
2006-03-01
期刊：
影响因子：
1.4
通讯作者：
Nolan, LK
中科院分区：
文献类型：
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作者：
Skyberg, JA;Logue, CM;Nolan, LK
本研究的目的是建立一种多重聚合酶链反应（PCR）技术，用于沙门氏菌的毒力基因分型。基因分型可以增强目前沙门氏菌的特征和分型方法。17个基因与沙门氏菌的入侵，菌毛生产，毒素生产，铁转运，和巨噬细胞内的生存有针对性的三个PCR反应。这些基因中的大多数是鼠模型中完整沙门氏菌毒力所需的，并且许多基因也位于沙门氏菌致病岛（PAI）上，并且与III型分泌系统（TTSS）相关。一旦使用阳性和阴性对照菌株的程序的成功得到验证，78沙门氏菌分离株的基因型在禽沙门氏菌病（主要是从生病，商业饲养的鸡和火鸡）和80沙门氏菌分离株明显健康的鸡或火鸡进行了比较。测试的17个基因中有11个（invA、orgA、prgH、WC、spaN [invJ]、sipB、sitC、pagC、msgA。spiA和iroN）。另一种（sopB）存在于所有分离的病禽和所有，但一个分离的健康鸟类。其余5个基因（lpfC、cdtB、sifA、pefA和spvB）在10%~ 90%的病禽和3.75%~ 90%的健康禽中被发现。两组分离物之间的这些基因的出现没有显着差异。这些结果表明，这些毒力基因，可能是PAI和TTSS与它们相关联，广泛分布在沙门氏菌分离株的鸟类，无论其来源的主机是否已被确定为沙门氏菌病。
The purpose of this study was to develop a multiplex polymerase chain reaction (PCR) protocol useful in the virulence genotyping of Salmonella spp. with the idea that genotyping could augment current Salmonella characterization and typing methods. Seventeen genes associated with Salmonella invasion, fimbrial production, toxin production, iron transport, and intramacrophage survival were targeted by three PCR reactions. Most of these genes are required for full Salmonella virulence in a murine model, and many are also located oil Salmonella pathogenicity islands (PAIs) and are associated with type III secretion systems (TTSSs). Once the success of procedures that used positive and negative control strains was verified, the genotypes of 78 Salmonella isolates incriminated in avian salmonellosis (primarily from sick, commercially reared chickens and turkeys) and 80 Salmonella isolates from apparently healthy chickens or turkeys were compared. Eleven of the 17 genes tested (invA, orgA, prgH, WC, spaN [invJ], sipB, sitC, pagC, msgA. spiA, and iroN) were found in all of the isolates. Another (sopB) was present in all isolates from sick birds and all but one isolate from healthy birds. The remaining five genes (lpfC, cdtB, sifA, pefA, and spvB) were found in 10%-90% of the isolates from sick birds and 3.75%-90% of the healthy birds. No significant differences in the Occurrence of these genes between the two groups of isolates were detected. These results Suggest that these virulence genes, and presumably the PAIs and TTSSs with which they are associated, are widely distributed among Salmonella isolates of birds, regardless of whether their hosts of origin have been identified as having salmonellosis.