Intestinal pathogenic Escherichia coli are the most common cause of diarrhoea and septicaemia in piglets, cattle and small ruminants, resulting in significant economic losses.
While most E. coli are harmless commensals which are part of the natural gastrointestinal flora of warm-blooded animals, some subsets, like the intestinal pathogenic E. coli, have acquired specific virulence-associated genes which enable them to be pathogenic in target hosts.
Virulence factors, responsible for pathogenicity, are fimbriae and adhesins as well as different toxins.
Detection and differentiation of virulence-associated genes of E. coli byusing Kylt® Multiplex Real-Time PCR systems enables determinationof potential virulence of E. coli isolates and allows to distinguishbetween different pathotypes (Table 1). Since these pathotypescause different symptoms in the host, the analysis of E. coli isolateswith Kylt® Multiplex Real-Time PCRs contributes to a differentiateddiagnosis.
| Pathotypes | Fimbriae and adhesins | Toxins | Kylt® Multiplex Real-Time PCR Kits covering fimbriae/adhesins and/or toxin |
|---|---|---|---|
| Enteropathogenic E. coli (EPEC) | Intimin (eae) | EAST | Kylt® E. coli eae, EAST |
| Enterotoxigenic E. coli (ETEC) | F4, F5, F6, F17, F18, F41, FimA/FimH, AIDA, paa | STI, STII, EAST, LTI, Stx2e, | Kylt® E. coli F4,F5,F6 Kylt® E. coli FimA, FimH, F41 Kylt® E. coli Sta, Stb, LT Kylt® E. coli F18, F41, Stx2e Kylt® E. coli F5, F17, F41 |
| Edema disease E. coli (EDEC) | F18, AIDA | Stx2e, EAST | Kylt® E. coli F18, F41, Stx2e Kylt® E. coli EAST, AIDA, paa |
| Shiga-toxin forming E. coli (STEC) | Intimin (eae) FimA, FimH | Stx1, Stx2, Stx2e | Kylt® E. coli Stx1, Stx2, eae (Feed & Food safety) |
Not all virulence-associated genes of E. coli are of clinical relevance in the different animal species. Some virulence factors are more likely to be found in E. coli strains isolated from swine, others are typical for E. coli isolated from cattle or small ruminants.
An overview which virulence factor could be of interest for clinical situations in different livestock animals can be found in table 2.
| Multiplex | Art. No 100 rxn | Art. No 25 rxn | Suckling piglets | weaners | ruminants | feed and food samples |
|---|---|---|---|---|---|---|
| Kylt® E. coli Sta, Stb, LT | 31706 | 31707 | x | x | x | |
| Kylt® E. coli F4, F5, F6 | 31710 | 31711 | x | x | ||
| Kylt® E. coli EAST, AIDA, paa | 31714 | 31715 | x | |||
| Kylt® E. coli FimA, FimH, F41 | 31718 | 31719 | x | |||
| Kylt® E. coli F18, F4, Stx2e | 31722 | 31723 | x | |||
| Kylt® E. coli F5, F17, F41 | 31726 | 31727 | x | |||
| Kylt® E. coli eae, EAST | 31730 | 31731 | x | |||
| Kylt® E.coli Stx1, Stx2, eae | 31734 | 31735 | x |
of E. coli by using Kylt® Multiplex Real-Time PCR
Suitable sample material:
Typing by Kylt® Multiplex Real-Time PCR systems should be
performed with individual isolates of E. coli collected from cultural processes of sample material derived from swine, cattle and small ruminants or feed and food samples.
Contents of the kits:
Kylt® Multiplex Real-Time PCR Kits comprise all reagents and controls for subsequent detection of E. coli virulence-associated genes, including a ready-to-use Reaction Mix for amplification as well as the Positive and Negative Controls.
Advantages
SPEED: Results can be achieved within 2-3 hours. The Real-Time PCR reaction itself takes less than 1.5 hours.
RELIABILITY: The Positive Control monitors the specificity and efficiency of the reagents and of the reaction itself. The Negative Control indicates absence of contaminations.
ACCURACY: The Internal Amplification Control is included in the Reaction Mix in an exact copy number; it is co-amplified in every single reaction to detect possible inhibitory effects of the DNA preparation and to verify true negative results.
The Kylt® Multiplex Real-Time PCR kits can be used on all commercially available Real-Time PCR thermal cyclers detecting the fluorescent dyes FAM, HEX, Cy5 and Texas Red.
Due to identical temperature profiles all Kylt® Multiplex Real-Time PCRs can be combined in one run as well as with other Kylt® Real-Time PCRs.