scholarly journals Molecular Characterization of Carbapenemase-Producing Escherichia coli and Salmonella in children with diarrhea in rural Burkina Faso

2020 ◽  
Author(s):  
Rene DEMBELE ◽  
Issiaka Soulama ◽  
Wendpoulomdé A. D. Kaboré ◽  
Ali Konaté ◽  
Assèta Kagambèga ◽  
...  
Keyword(s):  
Burkina Faso ◽  
Treatment Options ◽  
Diffusion Method ◽  
Chain Reaction ◽  
Disk Diffusion Method ◽  
Polymerase Chain ◽  
Pcr Assays ◽  
Resistance Rates ◽  
Encoding Genes

Abstract Background: In recent years, carbapenemase-producing Enterobacterales (CPE) resistance to antibiotics has dramatically increased leading to limitations of their treatment options. In the present study, we investigated the occurrence of carbapenemase-producing Escherichia coli and Salmonella in rural Burkina Faso. Methods: Salmonella isolates were serotyped according to the Kauffman White scheme. Diarrheagenic Escherichia coli (DEC) strains was identified using 16-plex Polymerase Chain Reaction (PCR), whereas antibiotic susceptibility was realized using the disk diffusion method. Furthermore, multiplex PCR assays were carried out using oligonucleotides to detect the presence of genes of the blaKPC, blaVIM, blaIMP, blaTEM, blaSHV, blaOXA and blaCTX-M types in all E. coli and Salmonella strains.Results: The study highlighted high resistance rates of the identified bacteria to common antibiotics. Likewise, two strains of E. coli were imipenem resistant with carbapenemase-encoding genes. The genes detected were Klebsiella pneumoniae carbapenemase (KPC), Verona integrin-encoded metallo-β-lactamase (VIM) and Imipenemase (IMP-2) reaching a rate of 40% each in E. coli strains. However, no Salmonella carbapenemases blaKPC, blaVIM or blaIMP were detected.Conclusions: This study showed that for a real-time infection control and prompt application of antimicrobial chemotherapy, characterization of carbapenemase-producing Enterobacterales in patients is crucial.

scholarly journals Molecular Characterization of Carbapenemase-Producing Enterobacterales in Children with Diarrhea in Rural Burkina Faso

2021 ◽  
Vol 11 (1) ◽  
pp. 84-92
Author(s):  
René Dembélé ◽  
Issiaka Soulama ◽  
Wendpoulomdé Aimé Désiré Kaboré ◽  
Ali Konaté ◽  
Assèta Kagambèga ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Burkina Faso ◽  
Treatment Options ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Klebsiella Pneumoniae Carbapenemase ◽  
Resistance Rates ◽  
Encoding Genes

Background and objective: In recent years, carbapenemase-producing Enterobacterales (CPE) resistance to antibiotics has dramatically increased leading to limitations of their treatment options. In the present study, we investigated the occurrence of carbapenemase-producing Escherichia coli and Salmonella in rural Burkina Faso.Materials and methods: Salmonella isolates were serotyped according to the Kauffman White scheme. Diarrheagenic Escherichia coli (DEC) strains was identified using 16-plex Polymerase Chain Reaction (PCR), whereas antibiotic susceptibility was realized using the disk diffusion method. Furthermore, multiplex PCR assays were carried out using oligonucleotides to detect the presence of genes of the blaKPC, blaVIM, blaIMP, blaTEM, blaSHV, blaOXA and blaCTX-M types in all E. coli and Salmonella strains.Results: The study highlighted high resistance rates of the identified bacteria to common antibiotics. Likewise, two strains of E. coli were imipenem resistant with carbapenemase-encoding genes. The genes detected were Klebsiella pneumoniae carbapenemase (KPC), Verona integrin-encoded metallo-β-lactamase (VIM) and Imipenemase (IMP-2) reaching a rate of 40% each in E. coli strains. However, no Salmonella carbapenemases blaKPC, blaVIM or blaIMP were detected.Conclusion: This study showed that for a real-time infection control and prompt application of antimicrobial chemotherapy, characterization of carbapenemase-producing Enterobacterales in patients is crucial.Keywords: Antibiotics, Carbapenemase-Producing Enterobacterales, children, Burkina Faso

scholarly journals Molecular Characterization of Carbapenemase-Producing Escherichia coli and Salmonella in children with diarrhea in rural Burkina Faso.

2020 ◽  
Author(s):  
Rene DEMBELE ◽  
Ali Konaté ◽  
Issiaka Soulama ◽  
Wendpoulomdé A. D. Kaboré ◽  
Assèta Kagambèga ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Burkina Faso ◽  
Treatment Options ◽  
Diffusion Method ◽  
Bacterial Strains ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Klebsiella Pneumoniae Carbapenemase ◽  
Encoding Genes

Abstract Background In recent years, Carbapenemase-producing Enterobacteriaceae (CPE) resistance to antibiotics has dramatically increased leading to limitations of their treatment options. In the present study, we investigated the occurrence of carbapenemase-producing Escherichia coli and Salmonella in rural Burkina Faso, using bacterial strains obtained from previous studies. Results Diarrheagenic Escherichia coli (DEC) strains was identified using 16-plex Polymerase Chain Reaction (PCR), whereas antibiotic susceptibility was realized using the disk diffusion method. Furthermore, multiplex PCR assays were used to characterize bla KPC, bla VIM and bla IMP genes in carbapenemase-producing E . coli and Salmonella . The study highlighted high resistance rates of the identified bacteria to common antibiotics. Likewise, two strains of E . coli were imipenem resistant with Carbapenemase-encoding genes. The genes detected were Klebsiella pneumoniae carbapenemase (KPC), Verona integrin-encoded metallo-β-lactamase (VIM) and Imipenemase (IMP-2) reaching a rate of 40% each. However, no Carbapenemase-encoding genes were detected in Salmonella isolates. Conclusions This study showed that for a real-time infection control and prompt application of antimicrobial chemotherapy, characterization of carbapenemase-producing Enterobacteriaceae in patients is crucial.

scholarly journals Occurrence of toxigenic Escherichia coli in raw milk cheese in Brazil

2007 ◽  
Vol 59 (2) ◽  
pp. 508-512 ◽  
Author(s):  
B.R. Paneto ◽  
R.P. Schocken-Iturrino ◽  
C. Macedo ◽  
E. Santo ◽  
J.M. Marin
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Antimicrobial Agents ◽  
Raw Milk ◽  
Diffusion Method ◽  
Chain Reaction ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Polymerase Chain ◽  
Raw Milk Cheese

The occurrence of toxigenic Escherichia coli in raw milk cheese was surveyed in Middle Western Brazil. Fifty samples of cheese from different supermarkets were analyzed for E.coli. The isolates were serotyped and screened for the presence of verotoxigenic E. coli (VTEC) and enterotoxigenic E. coli (ETEC) by Polymerase Chain Reaction (PCR). The susceptibility to thirteen antimicrobial agents was evaluated by the disk diffusion method. E.coli were recovered from 48 (96.0%) of the samples. The serogroups identified were O125 (6.0%), O111 (4.0%), O55 (2.0%) and O119 (2.0%). Three (6.0%) and 1(2.0%) of the E.coli isolates were VTEC and ETEC, respectively. Most frequent resistance was observed to the following antimicrobials: cephalothin (60.0%), nalidixic acid (40.0%), doxycyclin (33.0%), tetracycline (31.0%) and ampicillin (29.0%).

scholarly journals Detection of plasmid-mediated qnr genes among the quinolone-resistant Escherichia coli isolates in Iran

2014 ◽  
Vol 8 (07) ◽  
pp. 818-822 ◽  
Author(s):  
Farzaneh Firoozeh ◽  
Mohammad Zibaei ◽  
Younes Soleimani-Asl
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract ◽  
Urinary Tract Infections ◽  
Quinolone Resistance ◽  
Diffusion Method ◽  
Chain Reaction ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Polymerase Chain ◽  
Tract Infections

Introduction: Plasmid-mediated quinolone resistance, which complicates treatment, has been increasingly identified in Escherichia coli isolates worldwide. The purpose of this study was to identify the plasmid-mediated qnrA and qnrB genes among the quinolone-resistant Escherichia coli isolated from urinary tract infections in Iran. Methodology: A total of 140 Escherichia coli isolates were collected between March and October 2012 from urinary tract infections in Khorram Abad, Iran. All isolates were tested for quinoloe resistance using the disk diffusion method. Also, all quinolone-resistant isolates were screened for the presence of the qnrA and qnrB genes by polymerase chain reaction. Minimum inhibitory concentrations (MICs) of ciprofloxacin for the qnr-positive isolates were determined. Results: One hundred sixteen (82.8%) of 140 Escherichia coli isolates were nalidixic acid-resistant; among them, 14 (12.1%) and 9 (7.8%) were qnrA and qnrB-positive, respectively. Two quinolone-resistant isolates harbored both qnrA and qnrB. Among 63 ciprofloxacin-resistant isolates, 14 (22.2%) and 9 (14.3%) were found to carry qnrA and qnrB genes, respectively. The ciprofloxacin MIC range was 0.25–512 μg/mL for 23 qnr-positive Escherichia coli isolates, 18 of which had MICs values of 4–512 μg/mL. Conclusion: Our study shows that the frequency of plasmid-mediated quinolone resistance genes among E. coli isolates in Iran is high.

Ceftazidime – Avibactam susceptibility among carbapenem-resistant Enterobacterales in a pilot study in Turkey

2021 ◽  
Author(s):  
Hasan Selcuk Ozger ◽  
Ebru Evren ◽  
Serap Suzuk Yildiz ◽  
Cigdem Erol ◽  
Fatma Bayrakdar ◽  
...  
Keyword(s):  
Practical Method ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Drug Resistant ◽  
Chain Reaction ◽  
Disk Diffusion Method ◽  
Carbapenem Resistant ◽  
Therapeutic Choice ◽  
Polymerase Chain

AbstractThis study aimed to detect carbapenemase genes and to determine the in vitro susceptibility of Ceftazidime-Avibactam (CZA) in Enterobacterales isolates. Carbapenemase genes were detected by polymerase chain reaction. CZA sensitivity of isolates was evaluated with broth microdilution (BMD) and disk diffusion methods. A total of 318 carbapenem-resistant Enterobacterales isolates were included. Most of the isolates (n = 290, 91.2%) were identified as Klebsiella pneumoniae. The most common carbapenemase type was OXA-48 (n = 82, 27.6%). CZA susceptibility was evaluated in 84 isolates with OXA-48 and KPC carbapenemase activity. Both BMD and disk diffusion methods revealed that 95.2% of the isolates were sensitive to CZA; whereas, 4 (4.76%) isolates were resistant to CZA. Among colistin resistant isolates, 96.5% (n = 80) of them were susceptible to CZA. Our study demonstrated high in vitro efficacy of CZA in Enterobacterales isolates producing OXA-48 carbapenemase. High susceptibility rates against colistin resistant isolates which generally are also pan drug resistant, makes CZA a promising therapeutic choice for difficult-to-treat infections. Due to its high correlation with the BMD, disk diffusion method is a suitable and more practical method in detecting CZA in vitro activity.

scholarly journals Characterization of phytoplasmas related to 'Candidatus Phytoplasma asteris' subgroup rpI-L in Iran

2014 ◽  
Vol 54 (2) ◽  
pp. 199-203 ◽  
Author(s):  
Fereshteh Vali-Sichani ◽  
Masoud Bahar ◽  
Leila Zirak
Keyword(s):  
Fragment Length Polymorphism ◽  
Specific Primer ◽  
Sequence Analyses ◽  
Chain Reaction ◽  
Universal Primer ◽  
Disease Symptoms ◽  
Polymerase Chain ◽  
Pcr Assays ◽  
Niger Seed

Abstract In two of Iran's central provinces, several herbaceous plants showing phytoplasma disease symptoms were collected to detect 'Canididatus Phytoplasma asteris'-related phytoplasmas. Confirmation of an association of phytoplasmas with diseased plants was done using polymerase chain reaction (PCR) assays having the phytoplasma universal primer pairs P1/P7 followed by R16F2n/ R16R2 in nested PCR. Then, for detection of 'Ca. P. asteris', DNA samples were subjected to amplification of rp and tuf genes using specific primer pairs rp(I)F1A/rp(I)R1A and fTufAy/rTufAy, respectively. Restriction fragment length polymorphism or RFLP analyses of rp gene fragments using Tsp509I restriction enzyme as well as sequence analyses indicated that 'Ca. P. asteris'-related phytoplasmas associated with carrot, niger seed and scallion plants in these regions, belong to the rpI-L subgroup. This research is the first report of carrot, niger seed, and scallion infection with phytoplasmas belonging to the rpI-L subgroup.

scholarly journals Detection of Ampicillin Resistance Encoding Gene of Escherichia coli from Chickens in Bandung and Purwakarta

2019 ◽  
Vol 3 (1) ◽  
Author(s):  
Kuntum Khoirani ◽  
Agustin Indrawati ◽  
Surachmi Setiyaningsih
Keyword(s):  
Diffusion Method ◽  
Antibiotics Resistance ◽  
Test Results ◽  
Chain Reaction ◽  
Gene Encoding ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Polymerase Chain ◽  
Encoding Gene ◽  
Almost All

The purpose of this study was to test the resistance and to detect antibiotic resistence encoding gene in E. coli bacteria from chickens in Bandung and Purwakarta livestock. 18 E. coli isolates were tested for antibiotics resistance using the disk diffusion method. Isolates that were categorized as resistant and intermediate to antibiotics, then polymerase chain reaction was utilized to detect the resistent coding gene. The test results showed that all E. coli isolates from chickens in Bandung and Purwakarta were resistant to ampicillin (100%). E. coli isolates were still sensitive to chloramphenicol (11.1%) and gentamicin (22.2%). The gene encoding for ampC resistance from the test were in the amount of 77.7%. Sensitivity test results and detection of resistance coding gene showed that almost all isolates were resistant to ampicillin antibiotics and E. coli isolates were still sensitive to chlorampenicol and gentamicin. 

scholarly journals Evaluation of Two Phenotypic Methods for the Detection of Plasmid-Mediated AmpC β-Lactamases among Enterobacteriaceae Isolates

2021 ◽  
Author(s):  
Ronni Mol P ◽  
Khalid Mubarak Bindayna ◽  
Ganesan Shanthi
Keyword(s):  
Test Performance ◽  
Boronic Acid ◽  
Clinical Laboratory ◽  
Cost Effective ◽  
Tertiary Hospital ◽  
Diffusion Method ◽  
Chain Reaction ◽  
Disk Diffusion Method ◽  
Polymerase Chain ◽  
Phenotypic Methods

Abstract Objectives AmpC β-lactamases are cephalosporinases that confer resistance to cephalothin, cefazolin, cefoxitin, penicillin, and β-lactamase inhibitor-β-lactam combinations. Even though the AmpC resistance is reported, but the accurate occurrence of AmpC β-lactamases in Enterobacteriaceae members is still unknown. Techniques to identify AmpC producers are still evolving but not yet optimized for the clinical laboratory. Here we aimed to compare the test performance of two different phenotypic methods, that is inhibitor-based assay using boronic acid and disk approximation test for AmpC detection in Enterobacteriaceae isolates from a tertiary hospital microbiology laboratory. Materials and Methods The study includes 137 nonrepeat Enterobacteriaceae strains. Bacterial isolates, that yielded a zone diameter of less than 18 mm for cefoxitin by disk diffusion method were considered potential AmpC producers and further confirmed by phenotype methods—inhibitor-based assay using boronic acid and disk approximation test. A multiplex polymerase chain reaction was used to detect the most common plasmid-mediated AmpC genes: ACC, FOX, MOX, DHA, CIT, and EBC. Results Of the 137 clinical isolates, 58 (42.33%) were cefoxitin resistant, while 53.4 and 18.9% of the cefoxitin-resistant isolates were positive by inhibitor-based assay and disk approximation test. Multiplex PCR detected 42 (30.6%) isolates with AmpC genes. Of the 42 isolates, the inhibitor-based assay detected 25 (59.5%) isolates, while the disk approximation test detected nine (21.4%) isolates. Conclusion Our findings suggest that inhibitor-based assay using boronic acid can be used for the detection of the isolates that harbor AmpC β-lactamases. This method is cost-effective, simple to perform, and easy to interpret. Thus AmpC detection as a routine in clinical laboratories can help in appropriate therapeutic intervention and improved infection control.

scholarly journals Detection and Characterization of cry1Ac Transgene Construct in Bt Cotton: Multiple Polymerase Chain Reaction Approach

2007 ◽  
Vol 90 (6) ◽  
pp. 1517-1525 ◽  
Author(s):  
Chandra K Singh ◽  
Abhishek Ojha ◽  
Devendra N Kachru
Keyword(s):  
Polymerase Chain Reaction ◽  
Nested Pcr ◽  
Bt Cotton ◽  
Chain Reaction ◽  
Transcription Terminator ◽  
Npt Ii ◽  
Polymerase Chain ◽  
Pcr Assays ◽  
Multiple Polymerase Chain Reaction

Abstract To comply with international labeling regulations for genetically modified (GM) crops and food, and to enable proper identification of GM organisms (GMOs), effective methodologies and reliable approaches are needed. The spurious and unapproved GM planting has contributed to crop failures and commercial losses. To ensure effective and genuine GM cultivation, a methodology is needed to detect and identify the trait of interest and concurrently evaluate the structural and functional stability of the transgene insert. A multiple polymerase chain reaction (PCR) approach was developed for detection, identification, and gene stability confirmation of cry1Ac transgene construct in Bt cotton. As many as 9 samples of Bt cotton hybrid seeds comprising 3 approved Bt hybrids, MECH-12Bt, MECH-162Bt, MECH-184Bt, and a batch of 6 nonapproved Bt hybrids were tested. Initially, single standard PCR assays were run to amplify predominant GM DNA sequences (CaMV 35S promoter, nos terminator, and npt-II marker gene); a housekeeping gene, Gossypium hirsutum fiber-specific acyl carrier protein gene (acp1); a trait-specific transgene (cry1Ac); and a sequence of 7S 3 transcription terminator which specifically borders with 3 region of cry1Ac transgene cassette. The concurrent amplification of all sequences of the entire cassette was performed by 3 assays, duplex, triplex, and quadruplex multiplex PCR assays, under common assay conditions. The identity of amplicons was reconfirmed by restriction endonuclease digestion profile. The 2 distinct transgene cassettes, cry1Ac and npt-II, of the Bt cotton were amplified using the respective forward primer of promoter and reverse primer of terminator. The resultant amplicons were excised, eluted, and purified. The purified amplicons served as template for nested PCR assays. The nested PCR runs confirmed the transgene construct orientation and identity. The limit of detection as established by our assay for GM trait (cry1Ac) was 0.1. This approach can be adopted as a standard procedure for complete molecular characterization of Bt cotton. These assays will be of interest and use to importers, breeders, research laboratories, safety regulators, and food processors for detection of cry1Ac bearing GMOs.

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