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.

PathotypesFimbriae and adhesinsToxinsKylt® Multiplex Real-Time PCR
Kits covering fimbriae/adhesins and/or toxin
Enteropathogenic E. coli (EPEC)Intimin (eae)EASTKylt® E. coli eae, EAST
Enterotoxigenic E. coli (ETEC)F4, F5, F6, F17, F18, F41, FimA/FimH, AIDA, paaSTI, 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, AIDAStx2e, EASTKylt® E. coli F18, F41, Stx2e
Kylt® E. coli EAST, AIDA, paa
Shiga-toxin forming
E. coli (STEC)
Intimin (eae)
FimA, FimH
Stx1, Stx2,
Kylt® E. coli Stx1, Stx2, eae
(Feed & Food safety)
Table 1: Virulence factors for identification of E. coli pathotypes

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.

MultiplexArt. No
100 rxn
Art. No
25 rxn
weanersruminantsfeed and food
Kylt® E. coli Sta, Stb, LT3170631707xxx
Kylt® E. coli F4, F5, F63171031711xx
Kylt® E. coli EAST, AIDA, paa3171431715x
Kylt® E. coli FimA, FimH, F413171831719x
Kylt® E. coli F18, F4, Stx2e3172231723x
Kylt® E. coli F5, F17, F413172631727x
Kylt® E. coli eae, EAST3173031731x
Kylt® E.coli Stx1, Stx2, eae3173431735x
Table 2: Preferential host for taking sample material for detection of virulence-associated genes
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.

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.