Use of an enzyme-linked immunosorbent assay (ELISA) to measure the expression of the K88 fimbrial antigen by enterotoxigenic Escherichia coli (ETEC)

Payne, Dean; Moore, N. A.; Lambert, Peter A.

doi:10.1016/0022-1759(93)90169-8

Cited by 6 publications

(6 citation statements)

References 15 publications

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“…E. coli serotype 08:K87:K88ab:H19 was kindly supplied by R. Sellwood (Agricultural and Food Research Council, Institute of Animal Health, Compton, Newbury, Berkshire, United Kingdom). Bacteria were grown unshaken in nutrient broth (Oxoid Ltd., Basingstoke, United Kingdom) for 16 h at 37°C to maximize expression of K88 (16).…”

Section: Methodsmentioning

confidence: 99%

The K88 fimbrial adhesin of enterotoxigenic Escherichia coli binds to beta 1-linked galactosyl residues in glycosphingolipids

1993

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The K88 fimbrial adhesin enables certain strains of enterotoxigenic Escherichia coli to adhere to the porcine small intestine. In this study, the ability of the K88 adhesin to bind to glycosphingolipids was monitored by modified enzyme-linked immunosorbent assay and thin-layer chromatography overlay binding analysis. The binding of the K88 adhesin to glycosphingolipid-coated microtiter plates was saturable, with 50% maximal binding occurring with gangliotriaosylceramide, gangliotetraosylceramide, and lactosylceramide at 67 ± 21, 117 + 21, and 73 + 22 pM, respectively. Thin-layer chromatography overlay binding analysis demonstrated that serotype 08:K87:K88ab:H19 E. coli bound to hydroxylated galactosylceramide, gangliotriaosylceramide, gangliotetraosylceramide, and lactosylceramide but not to globotriaosylceramide, nonhydroxylated galactosylceramide, glucosides, glucosylceramide, or a mixture of ceramides. The K88 adhesin did not bind by either assay to globoside, the Forssman glycolipid, GM1, GM2, GM3, GDla, GDlb, GD2, GD3, GQlb, or GTlb. The binding pattern observed with the K88 adhesin suggests that o1-linked galactosyl residues are the minimum determinant required for binding, provided they are presented correctly. It is suggested that

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Section: Methodsmentioning

confidence: 99%

The K88 fimbrial adhesin of enterotoxigenic Escherichia coli binds to beta 1-linked galactosyl residues in glycosphingolipids

1993

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“…To this end, the bacteria have regulatory systems at their disposal that enable them to recognize the environmental circumstances and to adapt fimbriae biosynthesis according to these conditions. In the case of F4 fimbriae, expression is optimal at a temperature of 37°C, a pH ranging from 6.5 to 8.0 and at the end of the exponential growth [Jacobs et al, 1985;Mooi and de Graaf, 1985;van Verseveld et al, 1985;Blomberg et al, 1991;Payne et al, 1993]. This regulation of fimbrial expression is mediated by several factors like global regulators, local regulators, crosstalk between fimbriae and posttranscriptional mechanisms.…”

Section: Regulation Of F4 Expressionmentioning

confidence: 99%

F4 Fimbriae Expressed by Porcine Enterotoxigenic <i>Escherichia coli</i>, an Example of an Eccentric Fimbrial System?

Verdonck

Cox²,

Goddeeris³

2004

Microb Physiol

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An overwhelming number of infectious diseases in both humans and animals are initiated by bacterial adhesion to carbohydrate structures on a mucosal surface. Most bacterial pathogens mediate this adhesion by fimbriae or pili which contain an adhesive lectin subunit. The importance of fimbriae as virulence factors led to research elucidating the regulation of fimbrial expression and their molecular assembly process. This review provides an overview of the current knowledge of induction, expression and assembly of F4 (K88) fimbriae and discusses its unique as well as its identical characteristics compared to other intensively studied fimbriae or pili expressed by Escherichia coli.

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“…Clegg et al [19] in an ELISA to detect IgG against the CFAA antigen of ETEC from man established an optimal coating concentration of purified CFNI antigen of 1 pglml. On the other hand, Payne et al [38], without investigating the optimal coating antigen concentration in their ELISA assay for quantification of K88 antigen, described coating concentrations of purified K88 antigen as high as 10Opglml. In the present study, purified K88 antigen adsorbed to polystyrene plates at low concentrations, and antigen concentrations higher than the optimal decreased the OD values (Fig.…”

Section: Discussionmentioning

confidence: 99%

Development and evaluation of an ELISA to detect Escherichia coli K88 (F4) fimbrial antibody levels

Vázquez

González²,

Garabal³

et al. 1996

Journal of Medical Microbiology

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An enzyme-linked immunosorbent assay (ELISA) to determine IgG antibody levels against K88 (F4) fimbrial antigen from porcine enterotoxigenic Escherichia coli (ETEC) has been developed. The ELISA method was checked with serum samples obtained from rabbits and pigs, and the parameters affecting the method were also analysed. ELISA plates were optimally coated with K88 antigen 0.5 microgram/ml for testing rabbit antiserum or with 1.25 microgram/ml for testing pig serum. Optimal concentrations of H202 (0.5%) and orthophenylene-diamine (OPD) (0.125%) were chosen when a 10-min incubation period was used. The expression of antibody levels as enzyme-immunosorbent units (EIU) significantly decreased the variability of results between duplicate plates, when compared with the expression of results as direct OD values. ELISA-K88 applied to a field study with serum samples from 141 vaccinated and 52 unvaccinated sows was shown to be useful in differentiating between samples from vaccinated and unvaccinated animals.

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Use of an enzyme-linked immunosorbent assay (ELISA) to measure the expression of the K88 fimbrial antigen by enterotoxigenic Escherichia coli (ETEC)

Cited by 6 publications

References 15 publications

The K88 fimbrial adhesin of enterotoxigenic Escherichia coli binds to beta 1-linked galactosyl residues in glycosphingolipids

The K88 fimbrial adhesin of enterotoxigenic Escherichia coli binds to beta 1-linked galactosyl residues in glycosphingolipids

F4 Fimbriae Expressed by Porcine Enterotoxigenic <i>Escherichia coli</i>, an Example of an Eccentric Fimbrial System?

Development and evaluation of an ELISA to detect Escherichia coli K88 (F4) fimbrial antibody levels

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