Stanley J, Baquar N, Burnens A
Virus Reference Division, Central Public Health Laboratory, London, United Kingdom.
J Clin Microbiol. 1995 May;33(5):1206-11. doi: 10.1128/jcm.33.5.1206-1211.1995.
We describe a genotyping scheme for Salmonella panama. Defined probes specific for the 16S rRNA gene and the DNA insertion element IS200 were generated by PCR from S. panama and were used to probe genomic Southern blots made with enzymes selected to cut within and outside the probed sequences. Plasmid profiles were determined. The typeability and discriminatory power of the individual methods were compared. Ribotyping with 16S rRNA gene probe alone was slightly more discriminatory than phage typing, but unlike the latter, ribotyping was able to type all strains. IS200 profiling was the single most discriminatory method for S. panama, having an index of discrimination (D) of 0.8 and 100% typeability. Plasmid profiling, which had moderate discriminatory power but only 50% typeability, was valuable as an adjunct technique. The use of all three methods together or simply the combination of IS200 profiling with the two most discriminatory enzymes and plasmid profiling yielded a molecular typing scheme whose discriminatory power (D = 0.97) approached the maximum theoretical value. This should prove both useful and robust for epidemiological investigations of S. panama.
我们描述了一种用于巴拿马沙门氏菌的基因分型方案。通过PCR从巴拿马沙门氏菌中产生了针对16S rRNA基因和DNA插入元件IS200的特异性探针,并用于探测用选定酶切割的基因组Southern印迹,这些酶在被探测序列的内部和外部进行切割。测定了质粒图谱。比较了各个方法的分型能力和鉴别力。单独使用16S rRNA基因探针进行核糖体分型的鉴别力略高于噬菌体分型,但与噬菌体分型不同的是,核糖体分型能够对所有菌株进行分型。IS200谱型分析是对巴拿马沙门氏菌最具鉴别力的单一方法,鉴别指数(D)为0.8,分型能力为l00%。质粒图谱分析具有中等鉴别力,但分型能力仅为50%,作为辅助技术很有价值。同时使用这三种方法,或者简单地将IS200谱型分析与两种最具鉴别力的酶和质粒图谱分析相结合,产生了一种分子分型方案,其鉴别力(D = 0.97)接近最大理论值。这对于巴拿马沙门氏菌的流行病学调查应该是既有用又可靠的。