Comparison of PCR-based methods for typing Escherichia coli

Jonas, Daniel E; Spitzmüller, Bettina; Weist, Klaus; Rüden, H.; Daschner, Franz

doi:10.1046/j.1469-0691.2003.00661.x

Cited by 26 publications

(10 citation statements)

References 33 publications

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“…Our findings were coherent with other some studies (Fei et al, ; Wei et al, ; Mohapatra et al, ). On the other hand, the results were not in agreement with Jonas et al () who stated that RAPD and rep‐PCR methods were insufficient for the discrimination of E. coli strains also with Johnson and O’Bryan () who reported the inadequacy of single rep‐PCR methods in clear discrimination.…”

Section: Discussioncontrasting

confidence: 76%

Measurement of antibiotic resistance of microbial species in the Karasu River (Turkey) using molecular techniques

Avşar

2018

Water & Environment J

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The Karasu River (Sinop, Turkey) was sampled for bacteriological analyses and investigation of antibiotic resistance at the bacterial isolates. In the study, the physical properties and numbers of total aerobic (TA), total coliform (TC), faecal coliform (FC), faecal Streptococci (FS) and Escherichia coli in water samples were investigated. E. coli strains were subjected to antibiotic susceptibility testing, antibiotic resistant genes, virulence genes and plasmid DNA analysis and fingerprinting techniques. The bacteriological analysis results showed that the difference between TA, TC, FC and FS numbers (p > 0.05) was not significant for a total of four stations. The Multiple Antibiotic Resistance Index (MARI) values of tested strains were found in range from 0.4 to 0.7. The antibiotic resistance genes of strains; tetA (33.9%), blacym‐2 (12.5%) and cat1 (16.7%) were screened by PCR method. Out of a total of 57 E. coli isolates, 11 E. coli strains were positive for the eae (19.6%) gene, representing atypical EPEC, whilst only one strain was positive for the eae and stx1 (1.7%) gene, representing STEC. Present study has shown that the aquatic environments can be reservoirs of pathogenic E. coli strains. The study also indicated that PCR‐based techniques might especially be helpful in the rapid identification of multidrug‐resistance analysis and virulence genes of strains. Karasu stream, which is important in Sinop province, Turkey, was investigated for the first time in this study in terms of a molecular microbiological perspective.

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Section: Discussioncontrasting

confidence: 76%

Measurement of antibiotic resistance of microbial species in the Karasu River (Turkey) using molecular techniques

Avşar

2018

Water & Environment J

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“…To unequivocally prove identity of individual E. coli isolates, a highly discriminating typing method is needed. The use of antibiotic resistances profiles, PCRs targeting repetitive elements (rep‐PCRs) or multilocus sequence typing (MLST) (Rwego et al., ,b; Pesapane et al., ) failed to be as sensitive as PFGE analysis used in the current study (Jonas et al., ; McLellan et al., ). Although much more time and cost consuming than other methods, PFGE analysis' reproducibility is higher than that of other tests, for example rep‐PCR (Foley et al., ).…”

Section: Discussionmentioning

confidence: 72%

Experimental Evaluation of FaecalEscherichia coliand Hepatitis E Virus as Biological Indicators of Contacts Between Domestic Pigs and Eurasian Wild Boar

Barth

Geue

Hinsching

et al. 2015

Transbound Emerg Dis

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Domestic pigs and Eurasian wild boar (Sus scrofa) share several important viral and bacterial pathogens. Therefore, direct and indirect contacts between domestic pigs and wild boar present a risk of pathogen spillover and can lead to long-term perpetuation of infection. Biological indicators could be a powerful tool to understand and characterize contacts between wild boar and domestic pigs. Here, faecal Escherichia coli and Hepatitis E virus (HEV) were explored as potential biological indicators under experimental conditions. The data gained in our pilot study suggest that faecal E. coli can be used as biological indicator of contact between wild boar and domestic pig. For HEV, faecal transmission was also confirmed. However, molecular studies on full-genome basis did not reveal markers that would allow tracing of transmission direction. Based on these promising results, future field studies will especially target the practicability of E. coli microbiome molecular typing as surrogate of contacts at the wildlife-livestock interface.

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“…Several studies have recommended the usage of ERIC-PCR for E. coli strains obtained from different sources due to it is power of differentiation (Dhanashree and Shrikara, 2012 ;Diab and Al-urk, 2011;Maryam, 2016) In addition, several other PCR based typing methods have been developed based on the repeated DNA sequence elements distributed throughout the genome. These include randomly amplified polymorphic DNA and restriction fragment length polymorphism (Jonas et al, 2003).…”

Section: Enterobacterial Repetitive Intergenic Consensus (Eric)mentioning

confidence: 99%

Studies on avian pathogenic Escherichia coli in commercial broiler chicken in South East Queensland

Awawdeh¹,

Turni²,

Henning³

et al.

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Avian pathogenic Escherichia coli (APEC) is the causative agent of avian colibacillosis, a localised or systemic infection resulting in clinical diseases such as colisepticemia, chronic respiratory disease and swollen-head syndrome. Globally, avian colibacillosis is the leading cause of morbidity and mortality in poultry, and it has been associated with massive economic losses and welfare problems.This organism is of public health significance as APEC is communicable to humans. The diagnosis of avian colibacillosis relies on clinical signs, typical pathological lesions and culture of E. coli from affected tissue(s). Antimicrobial therapy is often used both for treatment and control. Previous overseas studies have characterised APEC and identified virulence genes (VGs) that can be used as molecular markers for the identification of APEC. Little is known about APEC in broiler chickens in Australia.The aim of this thesis was to gain a better understanding of the epidemiology of APEC in Australian broiler flocks and how factors including presence of VGs and phylogenetic group can improve the identification of this pathotype in Australia. Firstly, three faecal DNA extraction methods were evaluated. Faeces were collected from healthy chickens and chickens with colibacillosis from commercial broiler farms in South East Queensland (SEQ). The extracted DNA was screened by a pentaplex-PCR for five APEC-associated VGs (iroN, iutA, iss, hlyF and ompT). DNA extracted from E. coli isolates cultured from the cloaca and organs of the birds were screened using the same PCR.Repeated bead beating plus column elution was the preferred DNA extraction method, as it yielded good PCR quality and adequate quantity DNA. However, identifying APEC by direct detection of the five VGs from the faecal material was not feasible as all of these genes were also detected in all of the birds. However, the VGs were more commonly detected in E. coli isolates cultured from birds with colibacillosis.A cross-sectional study was performed to estimate APEC farm-level prevalence in healthy broiler chickens in SEQ and to identify potential risk factors associated with the carriage of APEC. At the farm-level, all of the 40 farms sampled were positive for APEC, that is at least one bird per farm carried APEC, while the within-farm prevalence was 63% (95% Confidence Interval: 55.8, 70.2).Higher APEC within-farm bird-level prevalence was significantly associated with the usage of well water as a source of drinking water, failure to disinfect the water line after each flock, farm visitors not showering before entering the shed, distances greater than 20 metres between the car park and the poultry shed and the presence of wild birds within 50 metres of the shed. Chlorinating the drinking water combined with automatic water filtration reduced within-farm bird-level APEC prevalence. Page | 3Therefore, based on the results concluded from the multivariable model, improving biosecurity and water treatments might reduce APEC prevalence, decrease the risk of co...

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Comparison of PCR-based methods for typing Escherichia coli

Cited by 26 publications

References 33 publications

Measurement of antibiotic resistance of microbial species in the Karasu River (Turkey) using molecular techniques

Measurement of antibiotic resistance of microbial species in the Karasu River (Turkey) using molecular techniques

Experimental Evaluation of FaecalEscherichia coliand Hepatitis E Virus as Biological Indicators of Contacts Between Domestic Pigs and Eurasian Wild Boar

Studies on avian pathogenic Escherichia coli in commercial broiler chicken in South East Queensland

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