scholarly journals Long-Term Carriage of Ciprofloxacin-Resistant Escherichia coli Isolates in High-Risk Nursing Home Residents

2016 ◽  
Vol 37 (4) ◽  
pp. 440-447 ◽  
Author(s):  
Miriam D. Ismail ◽  
Ting Luo ◽  
Sara McNamara ◽  
Bonnie Lansing ◽  
Evonne Koo ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
High Risk ◽  
Polymerase Chain Reaction Amplification ◽  
Nursing Home Residents ◽  
Multidrug Resistant ◽  
Chain Reaction ◽  
E Coli ◽  
Polymerase Chain ◽  
Gram Negative Organisms

BACKGROUNDRates of multidrug-resistant gram-negative organisms are surpassing those of methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci in nursing homes (NHs).OBJECTIVETo characterize the incidence and duration of carriage of ciprofloxacin-resistant Escherichia coli (CipREc) in NHs and identify those in the O25b-ST131 lineage.METHODSWe collected 227 CipREc isolates obtained by routine and regular surveillance of high-risk NH residents with indwelling devices. Repetitive element palindromic (REP)–polymerase chain reaction assay and multiplex polymerase chain reaction amplification for O25b-ST131 E. coli detection were performed using (GTG)5-primers and O25pabBspe and trpA2 primer pairs, respectively.RESULTSWe found a high period prevalence of CipREc colonization (21.5%), high rates of recolonization with the same strain following clearing (0.46 recolonizations/ person/ year), and an acquisition incidence of 1.05 cases/1,000 person-days. Almost three-quarters of colonized residents carried strains in the O25b-ST131 E. coli lineage. Compared with isolates not in the lineage, O25b-ST131 isolates were carried significantly longer (10 vs 3 months). We identified 18 different REP-types; 2 occurred in 55% of the residents colonized with CipREc, and in more than 1 NH. Duration of CipREc carriage varied by REP-type and averaged 6 months.CONCLUSIONCipREc occurred frequently in NH residents and is carried for long durations, and reacquisition following clearance is commonTrial registration. ClinicalTrials.gov identifier: NCT01062841.Infect. Control Hosp. Epidemiol. 2016;37(4):440–447

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 Multiplex Polymerase Chain Reaction Assay for Identification of Enterotoxigenic Escherichia Coli Strains

2001 ◽  
Vol 13 (4) ◽  
pp. 308-311 ◽  
Author(s):  
Jacek Osek
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Multiplex Polymerase Chain Reaction ◽  
Direct Determination ◽  
Enteric Bacteria ◽  
Enterotoxigenic Escherichia Coli ◽  
Chain Reaction ◽  
Gram Negative ◽  
E Coli ◽  
Polymerase Chain

A multiplex polymerase chain reaction (PCR) system was developed for identification of enterotoxigenic Escherichia coli (ETEC) strains and to differentiate them from other gram negative enteric bacteria. This test simultaneously amplifies heat-labile (LTI) and heat-stable (STI and STII) toxin sequences and the E. coli-specific universal stress protein ( uspA). The specificity of the method was validated by single PCR tests performed with the reference E. coli and non- E. coli strains and with bacteria isolated from pig feces. The multiplex PCR allowed the rapid and specific identification of enterotoxin-positive E. coli and may be used as a method for direct determination of ETEC and to differentiate them from other E. coli and gram-negative enteric isolates.

Polymerase Chain Reaction Assay for Detection of Escherichia coli O157:H7 and Escherichia coli O157:H−

2002 ◽  
Vol 65 (1) ◽  
pp. 5-11 ◽  
Author(s):  
TAKAHISA MIYAMOTO ◽  
NATSUKO ICHIOKA ◽  
CHIE SASAKI ◽  
HIROSHI KOBAYASHI ◽  
KEN-ICHI HONJOH ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Dna Sequence ◽  
Escherichia Coli O157 ◽  
Chain Reaction ◽  
Pcr Assay ◽  
E Coli ◽  
Pcr Product ◽  
Genomic Dnas ◽  
Polymerase Chain

The DNA band patterns generated by polymerase chain reaction (PCR) using the du2 primer and template DNAs from various strains of Escherichia coli and non–E. coli bacteria were compared. Among three to five prominent bands produced, the three bands at about 1.8, 2.7, and 5.0 kb were detected in all of the E. coli O157 strains tested. Some nonpathogenic E. coli and all pathogenic E. coli except E. coli O157 showed bands at 1.8 and 5.0 kb. It seems that the band at 2.7 kb is specific to E. coli O157. Sequence analysis of the 2.7-kb PCR product revealed the presence of a DNA sequence specific to E. coli O157:H− and E. coli O157:H7. Since the DNA sequence from base 15 to base 1008 of the PCR product seems to be specific to E. coli O157, a PCR assay was carried out with various bacterial genomic DNAs and O157-FHC1 and O157-FHC2 primers that amplified the region between base 23 and base 994 of the 2.7-kb PCR product. A single band at 970 bp was clearly detected in all of the strains of E. coli O157:H− and E. coli O157:H7 tested. However, no band was amplified from template DNAs from other bacteria, including both nonpathogenic and pathogenic E. coli except E. coli O157. All raw meats inoculated with E. coli O157:H7 at 3 × 100 to 3.5 × 102 CFU/25 g were positive both for our PCR assay after cultivation in mEC-N broth at 42°C for 18 h and for the conventional cultural method.

scholarly journals High Frequency of Diarrheagenic Escherichia coli in HIV-Infected Patients and Patients with Thalassemia in Kerman, Iran

2015 ◽  
Vol 16 (4) ◽  
pp. 353-358 ◽  
Author(s):  
Hesam Alizade ◽  
Hamid Sharifi ◽  
Zahedeh Naderi ◽  
Reza Ghanbarpour ◽  
Mehdi Bamorovat ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Statistical Analysis ◽  
High Frequency ◽  
Multiplex Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Fecal Samples ◽  
E Coli ◽  
Diarrheagenic Escherichia Coli ◽  
Polymerase Chain

This study was conducted on patients with thalassemia and HIV-infected patients to determine the frequency of diarrheagenic Escherichia coli in Kerman, Iran. We analyzed 68 and 49 E coli isolates isolated from healthy fecal samples of patients with thalassemia and HIV-infected patients, respectively. The E coli isolates were studied using a multiplex polymerase chain reaction to identify the enterotoxigenic E coli (ETEC), enterohemorrhagic E coli (EHEC), and enteropathogenic E coli (EPEC) groups. Statistical analysis was carried out to determine the correlation of diarrheagenic E coli between HIV-infected patients and patients with thalassemia using Stata 11.2 software. The frequency of having at least 1 diarrheagenic E coli was more common in patients with thalassemia (67.64%) than in HIV-infected patients (57.14%; P = .25), including ETEC (67.64% versus 57.14%), EHEC (33.82% versus 26.53%), and EPEC (19.11% versus 16.32%). The results of this study indicate that ETEC, EHEC, and EPEC pathotypes are widespread among diarrheagenic E coli isolates in patients with thalassemia and HIV-infected patients.

scholarly journals Use of Fluorescence Quantitative Polymerase Chain Reaction (PCR) for the Detection of Escherichia coli Adhesion to Pig Intestinal Epithelial Cells

2016 ◽  
Vol 19 (3) ◽  
pp. 619-625 ◽  
Author(s):  
C.H. Dai ◽  
L.N. Gan ◽  
W.U. Qin ◽  
C. Zi ◽  
G.Q. Zhu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Epithelial Cells ◽  
Intestinal Epithelial Cells ◽  
Quantitative Polymerase Chain Reaction ◽  
Relative Number ◽  
Intestinal Epithelial ◽  
Chain Reaction ◽  
E Coli ◽  
Polymerase Chain

AbstractAn efficient and accurate method to testEscherichia coli(E. coli) adhesion to intestinal epithelial cells will contribute to the study of bacterial pathogenesis and the function of genes that encode receptors related to adhesion. This study used the quantitative real-time polymerase chain reaction (qPCR) method. qPCR primers were designed from thePILINgene ofE. coliF18ab, F18ac, and K88ac, and the pig β-ACTINgene. Total deoxyribonucleic acid (DNA) fromE. coliand intestinal epithelial cells (IPEC-J2 cells) were used as templates for qPCR. The 2−ΔΔCtformula was used to calculate the relative number of bacteria in cultures of different areas. We found that the relative numbers of F18ab, F18ac, and K88ac that adhered to IPEC-J2 cells did not differ significantly in 6-, 12-, and 24-well culture plates. This finding indicated that there was no relationship between the relative adhesion number ofE. coliand the area of cells, so the method of qPCR could accurately test the relative number ofE. coli. This study provided a convenient and reliable testing method for experiments involvingE. coliadhesion, and also provided innovative ideas for similar detection methods.

Detection of enterotoxigenic Escherichia coli in water by polymerase chain reaction amplification and hybridization

1994 ◽  
Vol 40 (4) ◽  
pp. 243-249 ◽  
Author(s):  
Z. Tamanai-Shacoori ◽  
A. Jolivet-Gougeon ◽  
M. Pommepuy ◽  
M. Cormier ◽  
R. R. Colwell
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Amplification ◽  
Pcr Amplification ◽  
Environmental Water ◽  
Latex Agglutination ◽  
Enterotoxigenic Escherichia Coli ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chain Reaction ◽  
Polymerase Chain

Enterotoxigenic Escherichia coli was studied in waste water, river water, and seawater from six locations along the west coast of Normandy by using the polymerase chain reaction (PCR) to amplify the heat labile (LT) gene. Cellular DNA was extracted from centrifugation pellets and amplified using PCR. The PCR products were detected by gel electrophoresis and confirmed by hybridization assay, using an 850 base pair HindIII DNA fragment probe from pEWD299 conjugated to digoxigenin and specific for the LT gene. Results of the PCR amplification were compared with those of GM1 enzyme-linked immunosorbent assay, latex agglutination, and colony hybridization. The PCR method was found to be more precise and less time consuming, especially when compared with methods requiring culture of isolates for enumeration of enterotoxigenic E. coli in water.Key words: enterotoxigenic Escherichia coli, PCR, environmental water, digoxigenin.

scholarly journals Molecular Detection of Plasmid-Mediated Quinolone Resistance in Ciprofloxacin-Resistant Escherichia coli from Urine of Patients attending Garki Hospital, Abuja, Nigeria

2020 ◽  
Vol 1 (4) ◽  
Author(s):  
Moses Oghenaigah Eghieye ◽  
Istifanus Haruna Nkene ◽  
Rejoice Helma Abimiku ◽  
Yakubu Boyi Ngwai ◽  
Ibrahim Yahaya ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Urinary Tract ◽  
Molecular Detection ◽  
Quinolone Resistance ◽  
Chain Reaction ◽  
E Coli ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Tract Infections

Urinary tract infections (UTIs) caused by Escherichia coli (E. coli) is common worldwide; and its successful treatment using antibiotics is limited by acquisition of resistance by the bacteria. This study investigated the occurrence of plasmid-mediated quinolone resistance (PMQR) genes in ciprofloxacin-resistant E. coli from urine of patients with suspected cases of UTIs attending Garki Hospital Abuja (GHA), Nigeria. A total of 8 confirmed ciprofloxacin-resistant E. coli was screened for carriage of PMQR genes using polymerase chain reaction (PCR) method. The occurrences of the PMQR genes detected were in the order: aac-(6′)-Ib-cr (87.5%) > qnrB (50.0%) > qnrS (37.5%) > oqxAB (12.5%) > qnrA(0.0%). qnrB and qnrS did not exist alone, but in combination with other genes; aac-(6′)-Ib-crexisted both alone and in combination with others; the most prevalent patterns of existence were aac-(6′)-Ib-cr alone and aac-(6′)-Ib-cr + qnrB + qnrS at 25.0% each. This study has shown that the ciprofloxacin-resistant E. coli harbored aac-(6′)-Ib-cr, qnrB, qnrS and oqxAB PMQR genes, with aac-(6′)-Ib-cr being the most prevalent. The genes were present either alone or in combination with one another. This has implication for the clinical application of fluoroquinolones to treat UTI in the study location and environs. 

scholarly journals Screening of AmpC-/ESBL-producing Escherichia coli isolates from livestock for STEC/EHEC virulence genes

2021 ◽  
Vol 72 (3) ◽  
pp. 3147
Author(s):  
F PEHLIVANOGLU
Keyword(s):  
Public Health ◽  
Escherichia Coli ◽  
Virulence Genes ◽  
Multidrug Resistant ◽  
Beta Lactamase ◽  
Chain Reaction ◽  
Extended Spectrum Beta Lactamase ◽  
E Coli ◽  
Polymerase Chain ◽  
Flagellar Antigen

Livestock is an important reservoir of Shiga toxin-producing Escherichia coli and enterohemorrhagic E. coli (STEC/EHEC) strains and acts as a significant source of transmission to humans. In addition to the virulence of STEC/EHEC isolates, antibiotic resistance is also an escalating problem in these bacteria and increases the risk to public health. Therefore, the present study aimed to explore E. coli O157:H7 serotype and STEC/EHEC virulence genes in AmpC- and extended-spectrum beta-lactamase (ESBL)-producing E. coli isolates from cattle, chicken and sheep. A total of 61 confirmed AmpC- or ESBL-producing E. coli isolates were screened for the virulence genes (stx1, stx2, eae, ehxA, espP, katP and saa) and E. coli O157 (rfbO157) and H7 (fliCH7) genes by polymerase chain reaction (PCR). None of the ESBL-producing E. coli was positive for these genes, but six multidrug-resistant AmpC-producing E. coli were positive for the fliCH7 gene only. When considering the function of the H7 flagellar antigen of E. coli, it may be concluded that the development of ESBL/AmpC beta-lactamase production in the E. coli isolates with H7 flagella, which reside in the chicken intestine, may be potentially important for public health regarding both virulence and antimicrobial resistance.

A Multiplex Polymerase Chain Reaction Assay for Rapid Detection and Identification of Escherichia coli O157:H7 in Foods and Bovine Feces†

2000 ◽  
Vol 63 (8) ◽  
pp. 1032-1037 ◽  
Author(s):  
PINA M. FRATAMICO ◽  
LORI K. BAGI ◽  
TIZIANA PEPE
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Multiplex Pcr ◽  
Multiplex Polymerase Chain Reaction ◽  
Ground Beef ◽  
Escherichia Coli O157 ◽  
Chain Reaction ◽  
E Coli ◽  
Polymerase Chain ◽  
Bovine Feces

A multiplex polymerase chain reaction (PCR) assay was designed to simplify detection of Escherichia coli O157:H7 and to identify the H serogroup and the type of Shiga toxin produced by this bacterium. Primers for a plasmid-encoded hemolysin gene (hly933), and chromosomal flagella (fliCh7; flagellar structural gene of H7 serogroup), Shiga toxins (stx1, stx2), and attaching and effacing (eaeA) genes were used in a multiplex PCR for coamplification of the corresponding DNA sequences from enterohemorrhagic E. coli (EHEC) O157:H7. Enrichment cultures of ground beef, blue cheese, mussels, alfalfa sprouts, and bovine feces, artificially inoculated with various levels of E. coli O157:H7 strain 933, were subjected to a simple DNA extraction step prior to the PCR, and the resulting amplification products were analyzed by agarose gel electrophoresis. Sensitivity of the assay was ≤1 CFU/g of food or bovine feces (initial inoculum level), and results could be obtained within 24 h. Similar detection levels were obtained with ground beef samples that underwent enrichment culturing immediately after inoculation and samples that were frozen or refrigerated prior to enrichment. The multiplex PCR facilitates detection of E. coli O157:H7 and can reduce the time required for confirmation of isolates by up to 3 to 4 days.

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