Monospecific and bispecific antibodies against E. coli O157 for diagnostics

Guttikonda, Sujatha; Tang, X. L.; Yang, Bozka M.; Armstrong, Glen D.; Suresh, Mavanur R.

doi:10.1016/j.jim.2007.06.010

Cited by 18 publications

(14 citation statements)

References 23 publications

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“…Use of the ethanol precipitation of polysaccharides and of the phenol-chloroform extraction procedures described above was required for detection of as few as 7 CFU/liter of E. coli O157:H7 bacteria in highly turbid river water. This detection limit for E. coli O157:H7 bacteria in river water samples represents an improvement of several orders of magnitude over those of previous methods for water analysis (7,8,12). Direct PCR analysis of DNA was able to detect 2 ϫ 10 5 CFU/liter of E. coli O157:H7 bacteria in ground water (8).…”

Section: Resultsmentioning

confidence: 82%

“…The combination of this one-step capture-extraction method with the RT-PCR electronic DNA microarray detection technique was demonstrated to be capable of detecting 3 to 4 CFU of E. coli O157:H7 in 1 liter of tap water without cultural enrichment. Compared to the detection limits of 1,000 CFU/liter in drinking water (with DNA as a target) after 21 h of culture enrichment (7) and of 10 5 CFU/ liter (based on antibody determinations) in tap water (12), our method represents a substantial improvement for specific detection of viable E. coli O157:H7 bacteria. This method needs only minimum sample preparation and does not require culture enrichment.…”

Section: Resultsmentioning

confidence: 96%

“…As few as 6,000 CFU/liter in surface water could be detected using 21 h of enrichment and real-time PCR (with DNA as a target) (7). A sandwich enzyme-linked immunosorbent assay was able to detect 7.5 ϫ 10 5 CFU/liter in lake water (12). Detection of VBNC E. coli O157:H7 bacteria in river water.…”

Section: Resultsmentioning

confidence: 99%

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Detection of Viable but Nonculturable Escherichia coli O157:H7 Bacteria in Drinking Water and River Water

Liu

Gilchrist

Zhang

et al. 2008

Appl Environ Microbiol

116

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A sensitive method for specific detection of viable Escherichia coli O157:H7 cells, including viable but nonculturable (VBNC) cells, in water samples was developed. This method involved capture of the bacterial cells on a lowprotein-binding membrane and direct extraction and purification of RNA followed by reverse transcription-PCR and electronic microarray detection of the rfbE and fliC genes of E. coli O157:H7. It detected as few as 1 CFU of E. coli O157:H7 in diluted cultures, 3 to 4 CFU/liter in tap water, 7 CFU/liter in river water, and 50 VBNC cells in 1 liter of river water, demonstrating the best limit of detection reported to date for VBNC cells in environmental water samples.

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Section: Resultsmentioning

confidence: 82%

Section: Resultsmentioning

confidence: 96%

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Detection of Viable but Nonculturable Escherichia coli O157:H7 Bacteria in Drinking Water and River Water

Liu

Gilchrist

Zhang

et al. 2008

Appl Environ Microbiol

116

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“…Staphylococcus aureus (Guttikonda et al, 2007;Jung et al, 2003;McCarthy and Culloty, 2011;McClelland and Pinder, 1994 researchers use pure unconjugated primary antibodies to label target molecules. Thereafter, a range of commercially available secondary antibodies conjugated to a range of markers are used to reveal the labelled complex (primary antibody-antigen).…”

Section: O N O C Y T O G E N E S E S C H E R I C H I a C O L I O 1 mentioning

confidence: 99%

Application of Flow Cytometry to the Detection of Pathogenic Bacteria

Kennedy¹,

Wilkinson²

2017

Current Issues in Molecular Biology

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Outbreaks of infections have emphasized the necessity for rapid and economic detection methods for pathogens in samples ranging from those of clinical origin to food products during production and retail storage, and increasingly, in environmental samples. Flow cytometry (FCM) allows the rapid acquisition of multi-parametric data regarding cell populations within fluidised samples. However, the application of FCM to pathogen detection depends on the availability of specific fluorescent probes such as antibodies and RNA probes capable of detecting and isolating pathogens from these diverse samples. A particular issue for FCM methodology is the ability to recover and discriminate bacteria from the sample matrix which may pose a major technical hurdle towards accurate and sensitive analysis. This review article focuses on detection of pathogens using FCM in samples originating from food, water, environmental and clinical sources and outlines the current state of the art and potential future applications.

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“…It is important to note that low colony forming units (CFU) of E. coli O157:H7 present in contaminated food and water is generally sufficient to cause a severe infection, therefore having a sensitive method for detection during investigations of outbreaks is vital. A highly sensitive BsMAb-based immunodetection assay was developed by our laboratory for the detection of E. coli O157:H7 in water samples (Guttikonda et al 2007). E. coli O157:H7 whole bacteria and E. coli O157:H7 lipopolysaccharide (LPS) were used to first generate a monoclonal antibody, the hybridoma of which, was fused with anti-HRPO secreting monoclonal antibody hybridoma (Kreutz et al 1998;Suresh et al 1986b) to generate a quadroma producing BsMAb specific for both E. coli O157 and HRPO.…”

Section: Escherichia Coli O157:h7mentioning

confidence: 99%