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通过多重PCR快速、特异性检测腹泻和败血症性大肠杆菌菌株中与F17相关的菌毛蛋白和黏附素基因。

Rapid and specific detection of F17-related pilin and adhesin genes in diarrheic and septicemic Escherichia coli strains by multiplex PCR.

作者信息

Bertin Y, Martin C, Oswald E, Girardeau J P

机构信息

Laboratoire de Microbiologie, Centre de Recherche de Clermont-Ferrand-Theix, St Genès-Champanelle, France.

出版信息

J Clin Microbiol. 1996 Dec;34(12):2921-8. doi: 10.1128/jcm.34.12.2921-2928.1996.

Abstract

The F17-related adhesins are prevalent in Escherichia coli strains isolated from calves with diarrhea or septicemia and from lambs with nephropathy. The F17 family includes the F17a, F17b, F17c, and F111 fimbriae produced by bovine E. coli strains and the G agglutinin produced by human uropathogenic E. coli strains. An easy and inexpensive multiplex PCR method was developed to detect all the F17-related fimbriae and to identify four subtypes of structural subunit genes and two distinct subfamilies of adhesin genes by only two runs of amplification. A strict correlation was observed between the phenotypic assays and the multiplex PCR method when 166 pathogenic E. coli strains isolated from intestinal content of calves or lambs were tested. Genes encoding the F17c structural subunit and the subfamily II adhesins were prominent among the bovine and ovine isolates, and the capsule-like CS31A antigen was strictly associated with the F17c fimbriae. The F17b subtype fimbriae were prominent among the bovine isolates producing the CNF2 toxin, whereas the F17a subtype fimbriae were associated with the bovine isolates producing neither the CS31A antigen nor the CNF2 toxin. Five bacterial strains possessed two distinct and complete F17-related fimbrial gene clusters, and two of them produced two F17-related fimbriae at the bacterial cell surface. The related fimbrial gene clusters are probably organized in mosaic operons consisting of F17-related pilin and adhesin genes, and horizontal gene transfer may occur among E. coli strains isolated form different animal species.

摘要

F17相关黏附素在从患腹泻或败血症的犊牛以及患肾病的羔羊中分离出的大肠杆菌菌株中普遍存在。F17家族包括牛源大肠杆菌菌株产生的F17a、F17b、F17c和F111菌毛,以及人源致病性大肠杆菌菌株产生的G凝集素。开发了一种简便且廉价的多重PCR方法,通过仅两轮扩增就能检测所有F17相关菌毛,并鉴定结构亚基基因的四个亚型和黏附素基因的两个不同亚家族。当对从犊牛或羔羊肠道内容物中分离出的166株致病性大肠杆菌菌株进行检测时,在表型分析和多重PCR方法之间观察到了严格的相关性。编码F17c结构亚基和亚家族II黏附素的基因在牛和羊的分离株中很突出,并且类荚膜CS31A抗原与F17c菌毛严格相关。F17b亚型菌毛在产生CNF2毒素的牛分离株中很突出,而F17a亚型菌毛与既不产生CS31A抗原也不产生CNF2毒素的牛分离株相关。五株细菌菌株拥有两个不同且完整的F17相关菌毛基因簇,其中两株在细菌细胞表面产生两种F17相关菌毛。相关菌毛基因簇可能由F17相关菌毛蛋白和黏附素基因组成镶嵌操纵子,并且水平基因转移可能发生在从不同动物物种分离出的大肠杆菌菌株之间。

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