A colony blot immunoassay to detect enteroinvasive Escherichia coli and Shigella in water samples

Szakál, D.; Gadó, I.; Pal, Tibor

doi:10.1046/j.1365-2672.2001.01242.x

Cited by 10 publications

(9 citation statements)

References 25 publications

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“…The presence of faecal coliforms in water indicates contaminant per se, and may indicate that the sample is unsuitable for consumption, (Szakal et al, 2001). Ahmed et al, (2005) reported that the use of indicator bacteria such as faecal coliforms fc and Escherichia coli for the assessment of faecal pollution and possible water quality deterioration in various water sources is widely accepted concept in the world.…”

Section: Introductionmentioning

confidence: 99%

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Prevalence of <i>Escherichia coli</i> some public water sources in Gusau Municipal, North Western Nigeria

Garba¹,

Mohammed²,

Yakubu³

et al. 2011

Bayero J. Pure App. Sci.

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This study investigated the presence of Escherichia coli from some public water sources in Gusau municipal, northwestern Nigeria. This was done by determining the total coliform counts and the presence of Escherichia coli and its antibiotic susceptibility profile. A total of 180 well 60 tap and 60 packaged water samples were obtained from Gusau municipal on weekly basis over a period of seven month (August, 2006-Feb, 2007) covering part of rainy and dry seasons. Standard procedures were used for the identification of Escherichia coli and for estimating total coliform counts. All samples had coliform counts higher than the international standard recommended by the World Health Organization. Results of biochemical analysis of the samples showed that out of 63 confirmed Escherichia coli isolated, 41 (45.5%) were from well water, tap water had 14 (23.3%) while packaged water had 8(13.3%).The susceptibility profile of the isolates to nine antimicrobial agents indicated that majority of the isolates were highly susceptible to Chloramphenicol, Gentamycin, Perrfloxacin, Tarivid, Augmentin, Streptomycin, Sparfloxacin, and Ciprofloxacin, moderate susceptibility to Septrin and resistant to Amoxacilin were observed. None of the water samples met the WHO standards for drinking water and thus pose a serious health risk to its consumers and users if not properly treated.

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Section: Introductionmentioning

confidence: 99%

“…The role of food and water as vehicle for the transmission of this organism is well documented and represent a significant public health threat (Szakal et al, 2001). This study investigates the microbiological quality of water sources in Gusau municipal.…”

Section: Introductionmentioning

confidence: 99%

Prevalence of <i>Escherichia coli</i> some public water sources in Gusau Municipal, North Western Nigeria

Garba¹,

Mohammed²,

Yakubu³

et al. 2011

Bayero J. Pure App. Sci.

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“…A protein A-alkaline phosphatase conjugate solution (Sigma) was diluted 1:500 in 2% casein and added to the blots for 1 h. The blots were washed again in TBS and developed with Naphthol/Fast red solution in 50 mM Tris-HCl, pH 8.0 (Sigma). This protocol was adapted from protocols described by Szakal et al (48).…”

Section: Methodsmentioning

confidence: 99%

“…In this study, a simple colony blot technique, adapted from previous studies with Shigella (48), was used to demonstrate the surface expression of the CfaB and LTB proteins in SC608(pCFAI) and SC608(pC-FAI/LTB). This technique allows for the adsorption of extracellular proteins directly onto nitrocellulose for detection by immunoblotting.…”

Section: Methodsmentioning

confidence: 99%

Construction and Characterization of Bivalent Shigella flexneri 2a Vaccine Strains SC608(pCFAI) and SC608(pCFAI/LTB) That Express Antigens from Enterotoxigenic Escherichia coli

et al. 2005

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An invasive strain of Shigella flexneri 2a (SC608) has been developed as a vector for the expression and delivery of heterologous antigens. SC608 is an aspartate semialdehyde dehydrogenase (asd) derivative of SC602 (icsA iuc), a well-characterized live attenuated vaccine strain which has undergone several clinical trials in human volunteers. When administered orally at a single 10 4 (CFU) dose, SC602 is both immunogenic and efficacious against shigellosis. Using asd-based plasmid vectors, we designed SC608 to express the enterotoxigenic Escherichia coli (ETEC) fimbrial subunit CfaB (CFA/I structural subunit) alone or in combination with the E. coli B subunit of heat-labile enterotoxin (LTB). The expression of each heterologous protein in SC608 was verified by immunoblot analysis. Each strain was comparable to the parent strain, SC602, in a HeLa cell invasion assay. After intranasal immunizations of guinea pigs, serum and mucosal immune responses were detected against both Shigella lipopolysaccharide and heterologous ETEC antigens by enzyme-linked immunosorbent assay and ELISPOT analysis. All immunized animals were subsequently protected against a challenge with wild-type S. flexneri 2a in a keratoconjunctivitis Sereny test. Serum antibodies generated against LTB and CfaB demonstrated antitoxin and agglutination activities, respectively. These results suggest that CfaB and LTB expressed in SC608 retain important conformational epitopes that are required for the generation of antibodies that have functional activities. These initial experiments demonstrate that a fully invasive Shigella vaccine strain can be engineered to deliver antigens from other diarrheal pathogens.

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“…They can detect only 10 5 ± 10 6 cells/mL without pre-enrichment. Methods such as the colony blot immunoassay [3] need a very high concentration of the bacterial colonies to be present on the membranes for the evaluation of the immuno reaction. To overcome these problems, alternative immunoassay techniques are being developed that aim at reducing the analysis time, simplifying the analysis procedure, improving sensitivity (to over 2000 times higher sensitivity than ELISA), while allowing their application to field conditions.…”

Section: Introductionmentioning

confidence: 99%

Rapid Amperometric Immunoassay for Escherichia coli Based on Graphite Coated Nylon Membranes

Shah¹,

Chemburu²,

Wilkins³

et al. 2003

Electroanalysis

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An amperometric immunosensor for rapid detection of bacteria has been developed. A graphite-coated nylon membrane serves as a support for antibody immobilization and as a working electrode. The different parameters affecting sensor preparation such as, selection of the conductive material, pore size of the nylon membrane, number of coatings and the ratios of the graphite to the binder were optimized. A ™sandwich∫ immunoassay scheme was used and amperometric detection of captured bacteria was conducted by the electro-reduction of iodine formed as the product of a peroxidase catalyzed reaction. This approach has been used for detection of total Escherichia coli with a low detection limit of 40 cells/mL. The developed system can be easily adapted for rapid analysis of different pathogenic microorganisms.

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A colony blot immunoassay to detect enteroinvasive Escherichia coli and Shigella in water samples

Cited by 10 publications

References 25 publications

Prevalence of <i>Escherichia coli</i> some public water sources in Gusau Municipal, North Western Nigeria

Prevalence of <i>Escherichia coli</i> some public water sources in Gusau Municipal, North Western Nigeria

Construction and Characterization of Bivalent Shigella flexneri 2a Vaccine Strains SC608(pCFAI) and SC608(pCFAI/LTB) That Express Antigens from Enterotoxigenic Escherichia coli

Rapid Amperometric Immunoassay for Escherichia coli Based on Graphite Coated Nylon Membranes

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