通过直接PCR扩增hilA基因简单快速地检测沙门氏菌菌株。
Simple and rapid detection of Salmonella strains by direct PCR amplification of the hilA gene.
作者信息
Pathmanathan S G, Cardona-Castro N, Sánchez-Jiménez M M, Correa-Ochoa M M, Puthucheary S D, Thong K L
机构信息
Institute of Postgraduate Studies1, Department of Medical Microbiology, Faculty of Medicine4 and Institute of Biological Sciences5, University of Malaya, Kuala Lumpur, Malaysia 2Instituto Colombiano de Medicina Tropical, Sabaneta, Colombia 3Escuela de Bacteriología, Universidad de Antioquia, Medellín, Colombia.
出版信息
J Med Microbiol. 2003 Sep;52(Pt 9):773-776. doi: 10.1099/jmm.0.05188-0.
The suitability of a PCR procedure using a pair of primers targeting the hilA gene was evaluated as a means of detecting Salmonella species. A total of 33 Salmonella strains from 27 serovars and 15 non-Salmonella strains from eight different genera were included. PCR with all the Salmonella strains produced a 784 bp DNA fragment that was absent from all the non-Salmonella strains tested. The detection limit of the PCR was 100 pg with genomic DNA and 3 x 10(4) c.f.u. ml(-1) with serial dilutions of bacterial culture. An enrichment-PCR method was further developed to test the sensitivity of the hilA primers for the detection of Salmonella in faecal samples spiked with different concentrations of Salmonella choleraesuis subsp. choleraesuis serovar Typhimurium. The method described allowed the detection of Salmonella Typhimurium in faecal samples at a concentration of 3 x 10(2) c.f.u. ml(-1). In conclusion, the hilA primers are specific for Salmonella species and the PCR method presented may be suitable for the detection of Salmonella in faeces.
评估了使用一对靶向hilA基因的引物的PCR程序作为检测沙门氏菌属的一种方法。共纳入了来自27个血清型的33株沙门氏菌菌株和来自8个不同属的15株非沙门氏菌菌株。所有沙门氏菌菌株的PCR均产生了一个784 bp的DNA片段，而所有测试的非沙门氏菌菌株均未产生该片段。PCR对基因组DNA的检测限为100 pg，对细菌培养物系列稀释液的检测限为3×10⁴ c.f.u. ml⁻¹。进一步开发了一种富集PCR方法，以测试hilA引物对检测掺入不同浓度猪霍乱沙门氏菌亚种猪霍乱血清型鼠伤寒沙门氏菌的粪便样本中沙门氏菌的敏感性。所描述的方法能够检测粪便样本中浓度为3×10² c.f.u. ml⁻¹的鼠伤寒沙门氏菌。总之，hilA引物对沙门氏菌属具有特异性，所提出的PCR方法可能适用于检测粪便中的沙门氏菌。
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