Sayada C, Picard B, Elion J, Krishnamoorthy R
Laboratoire de Biochimie Génétique, Hôpital Robert Debré, Paris, France.
Mol Cell Probes. 1994 Jun;8(3):187-91. doi: 10.1006/mcpr.1994.1025.
The identification and differentiation of the two variants of the ail gene, ailA from the more virulent American serotypes (08, 013a, 13b, 018, 020, 021) and ailNA from the less virulent non-American serotypes (03, 04, 05, 06, 09, 027 and 07, 8) was studied in a panel of 32 Yersinia enterocolitica human pathogenic isolates. A 444 bp fragment corresponding to the ail gene was amplified using a PCR procedure in all tested strains. Subsequent digestion of the PCR product by Rsal and by HaeIII endonucleases, provide electrophoretic patterns that clearly discriminate ailA and ailNA variants. This non-radioactive and reliable procedure allows large clinical and epidemiological studies, and could be proposed to survey the spread of virulent clones.
在一组32株人致病性小肠结肠炎耶尔森氏菌分离株中,研究了ail基因的两种变体的鉴定和区分,即来自毒力更强的美国血清型(08、013a、13b、018、020、021)的ailA和来自毒力较弱的非美国血清型(03、04、05、06、09、027以及07、8)的ailNA。使用PCR程序在所有测试菌株中扩增出对应于ail基因的444 bp片段。随后用Rsal和HaeIII核酸内切酶对PCR产物进行消化,得到能清晰区分ailA和ailNA变体的电泳图谱。这种非放射性且可靠的程序适用于大规模临床和流行病学研究,可用于调查毒力克隆的传播情况。