Application of Molecular Diagnostic Techniques for the Detection of E. coli O157:H7: A Review

Tamerat, Nateneal; Muktar, Yimer

doi:10.4172/2157-7579.1000362

Cited by 6 publications

(7 citation statements)

References 43 publications

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Section: Resultsmentioning

confidence: 99%

“…Conventional methods for specific quantification and differentiation of microbial cells use either selective culturing media, which can take up to several days to distinguish a positive from a negative sample, or molecular biology techniques, which instead target mainly intracellular biomarkers (i.e., proteins, nucleic acids) and therefore require complex and time-consuming procedures for extraction, amplification, and revelation (i.e., immunolabeling, PCR). 29 A less explored strategy for cell sensing focuses instead on the extracellular complex array of macro/biomolecules expressed on bacterial cell walls (i.e., phospholipids, lipopolysaccharides). Such sensing strategy takes advantage of the chemical fingerprint of these complex moieties to generate a nonspecific but selective response relying on the differential binding affinities between different nanoprobes and bacterial cells, thus without the need of costly biological receptors (i.e., antibodies, peptides, and nucleic acids).…”

Section: ■ Introductionmentioning

confidence: 99%

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Low-Cost Strategy for the Development of a Rapid Electrochemical Assay for Bacteria Detection Based on AuAg Nanoshells

et al. 2018

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A low-cost strategy for the simple and rapid detection of bacterial cells in biological matrixes is presented herein. Escherichia coli and Salmonella typhimurium were chosen as model bacteria for the development of an electrochemical assay based on hollow AuAg nanoshells (NSs). By taking advantage of their electrocatalytic properties for the in situ generation of the electrochemical signal without the need of any other kind of reagent, substrate, or redox enzyme, high sensitivities (down to 10 2 CFU/mL) were achieved. Moreover, the recognition and discrimination of the model bacterial cells in the sample matrix was possible by relying solely on nonspecific affinity interactions between their cell walls and AuAg NSs surface, avoiding the use of expensive and fragile biological receptor. Compared to traditional, laboratorybased analytical tests available, this assay provides a promising proof-of-concept alternative that allows to obtain good sensitivities and selectivity in very short times in addition to the low cost.

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Section: Resultsmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 99%

Low-Cost Strategy for the Development of a Rapid Electrochemical Assay for Bacteria Detection Based on AuAg Nanoshells

et al. 2018

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“…A possible explanation is that the O157:H7 strain did not express characteristically or the antigenic protein got saturated by host antibodies. Recent advances in diagnostic techniques have made several protocols available for isolation and characterization of E. coli O157:H7 from food, fecal, and environmental samples (Deisingh & Thompson, 2004;Tamerat, Muktar, & Shiferaw, 2016). Further, to classify and elaborate the function of the main virulence genes, such as stx 1 and stx 2 , and genes encoding accessory STEC virulence factors, such as eaeA, hlyA, and flic H7 , different PCR assays have been described (Al-Ajmi et al, 2006;Gannon, King, Kim, & Thomas, 1992;Holland, Louie, Simor, & Louie, 2000;Kumar, Grover, & Kumar Batish, 2013;Pan, Chen, & Su, 2002;Sharma, 2006;Sharma & Dean-Nystrom, 2003).…”

Section: Discussionmentioning

confidence: 99%

“…E. coli serotype O157:H7 was discovered in 1982 (Riley et al, 1983), it has produced outbreaks worldwide and is ranked among the most studied foodborne pathogens (Adamu, Shamsul, Desa, & Khairani-Bejo, 2014;Bell et al, 1994;Ibrahim, 2015;Kieckens et al, 2015;Torso et al, 2015). Natural reservoirs of this pathogen include cattle, sheep, and goats, and the modes of transmission of the infections are animal to person, waterborne, foodborne, and person to person (Ateba & Mbewe, 2011;Ferens & Hovde, 2011).…”

Section: Introductionmentioning

confidence: 99%

Multiplex polymerase chain reaction detection of Shiga toxin genes and antibiotic sensitivity of Escherichia coli O157:H7 isolated from beef meat in Quetta, Pakistan

Samad

Abbas

Ahmad

et al. 2018

Journal of Food Safety

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The objective of this study was to conduct a qualitative analysis of raw beef meat sold in the city of Quetta, Pakistan for presence and drug sensitivity of the potentially pathogenic Escherichia coli strain O157:H7. The study used 200 raw beef meat samples collected from retail butcher shops. Conventional and rapid biochemical tests, latex agglutination and multiplex polymerase chain reaction (PCR) using primers designed for the rfb O157 and flic H7 genes were used to detect E. coli O157:H7. All O157:H7 isolates were also tested for Shiga toxin genes stx 1 and stx 2. The prevalence of E. coli O157:H7 in collected beef meat samples was 10%. Detection through PCR was found more sensitive than detection of O and H antigens. The quantity of E. coli O157:H7 isolates positive for Shiga toxins was 50% (20% for stx 1, 45% for stx 2 and 10% for both stx 1 and stx 2). Season wise variation showed highest E. coli O157:H7 prevalence during summer months. A further concern is that E. coli O157:H7 isolates were resistant to a range of common antibiotics. The results indicate an urgent need for applying proper food hygiene practices in the Quetta region to reduce incidence of foodborne diseases and they also emphasize the global problem of antimicrobial resistance. Practical applications E. coli O157:H7 is as a potentially threatening foodborne pathogen. A significant prevalence of E. coli O157:H7 detected in raw beef meat from retail outlets in the city of Quetta indicates an urgent need for applying proper food hygiene practices in the Quetta region to reduce the incidence of foodborne diseases. Furthermore, resistance of the E. coli O157:H7 isolates to a range of commonly used antibiotics emphasizes the global problem of antimicrobial resistance. The multiplex PCR method used here is a reliable, sensitive, and relatively rapid technique for detecting E. coli O157:H7 in food and environmental samples and important for ongoing surveillance to minimize contamination of raw meat products and associated cross contamination by E. coli O157:H7.

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“…The Multiple Antibiotic Resistance (MAR) index was calculated and interpreted as a/b, where "a" represents the number of antibiotics to which the isolate was resistant, and "b" represents the number of antibiotics to which the isolate was exposed [17]. Lysing was done by putting a colony of E. coli in 30 µL DNAse/RNAse free water and lysed at 99 • C for 30 min [18] in a thermocycler (peqSTAR 96X Universal gradient thermocycler, VWR, Darmstadt, Germany). The lysate was then used as the template for PCR amplification.…”

Section: Antimicrobial Susceptibility Of E Colimentioning

confidence: 99%

Antimicrobial Susceptibility and Molecular Characterization of Escherichia coli Recovered from Milk and Related Samples

et al. 2022

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There is a rising concern about illnesses resulting from milk consumption due to contamination by pathogenic microorganisms including Escherichia coli. This study examined the occurrence and antimicrobial susceptibility of E. coli isolated from cow milk and related samples. Furthermore, partial sequencing was done to ascertain the genetic relatedness and possible cross contamination among the samples. In all, 250 samples, that is, 50 each of raw milk, cow teat, milkers’ hands, milking utensils, and fecal matter of cows, were cultured for the identification of E. coli. E. coli was detected in 101/250 samples (40.4%). Milk and fecal samples recorded the highest percentages of 68.0% and 66.0%, respectively. Forty-two (42) E. coli strains examined for antimicrobial resistance showed an overall 25.5% resistance, 15.0% intermediate resistance, and 59.5% susceptibility. The isolates had a high level of resistance to teicoplanin (100.0%), but were susceptible to chloramphenicol (95.2%) and azithromycin (92.9%). The Multiple Antibiotic Resistance (MAR) index pattern ranged from 0.1 to 0.5, and 40.5% exhibited multiple drug resistance. The E. coli strains formed 11 haplotypes, and a phylogenic tree analysis showed relatedness among the isolates in other African countries. This observation is an indication of cross contamination among the milk and its related samples.

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Application of Molecular Diagnostic Techniques for the Detection of E. coli O157:H7: A Review

Cited by 6 publications

References 43 publications

Low-Cost Strategy for the Development of a Rapid Electrochemical Assay for Bacteria Detection Based on AuAg Nanoshells

Low-Cost Strategy for the Development of a Rapid Electrochemical Assay for Bacteria Detection Based on AuAg Nanoshells

Multiplex polymerase chain reaction detection of Shiga toxin genes and antibiotic sensitivity of Escherichia coli O157:H7 isolated from beef meat in Quetta, Pakistan

Antimicrobial Susceptibility and Molecular Characterization of Escherichia coli Recovered from Milk and Related Samples

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