scholarly journals Antibiotic Resistance Pattern and Detection of Enterotoxigenic and Enteroaggregative Strain of Escherichia coli among Foreign Tourist with Traveler Diarrhea in Bali using Uniplex Polymerase Chain Reaction

2020 ◽  
Vol 8 (A) ◽  
pp. 583-588
Author(s):  
I Dewa Made Sukrama ◽  
Anak Agung Wiradewi Lestari ◽  
Made Agus Hendrayana ◽  
I Ketut Agus Somia
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Significant Proportion ◽  
Resistance Pattern ◽  
Beta Lactam ◽  
First Line ◽  
Chain Reaction ◽  
Sensitivity Testing ◽  
E Coli ◽  
Polymerase Chain

BACKGROUND: As one of the major tourist destinations in Southeast Asia, Bali received millions of foreign tourists each year. Diarrhea consistently placed as the most often experienced health problem among travelers. Traveler diarrhea has various etiologies. The most common was Escherichia coli. The existence of several types of E. coli that are resistant to several antibiotics causes the selection of antibiotics is crucial. AIM: This preliminary study aims to understand the pattern of antibiotics sensitivity and to detect the presence of enterotoxigenic and enteroaggregative strains of E. coli from fecal samples of foreign tourists with traveler’s diarrhea in Denpasar, Bali. METHODS: A culture examination was carried out to obtain E. coli bacterial colonies. Disk diffusion Kirby–Bauer was carried out for antibiotic sensitivity testing. The confirmed colonies were tested against several common antibiotics, including the recommended first line (ciprofloxacin and azithromycin). Uniplex polymerase chain reaction (PCR) using specific primers conducted to detect the enterotoxigenic E. coli (ETEC) (elt and estA2-4) and enteroaggregative E. coli (EAEC) (CVD432) strains. RESULTS: Among 48 stool culture, 14 (29.2%) were identified as E. coli colonies. All samples were still sensitive to the antibiotics meropenem, ceftazidime, and cefixime. Despite majority of the samples (78.6%) still sensitive to ciprofloxacin, large proportion of the samples have developed resistance against the other commonly used antibiotics, doxycycline (70.4%) and azithromycin (57.1%). PCR showed that 3 (21.4%) samples shown positive for CVD432 gene, 2 (14.3%) samples positive for the elt gene, and all negative for the estA2-4 gene. CONCLUSION: An only small proportion of E. coli was EAEC or ETEC strain. Although most E. coli still sensitive to beta-lactam antibiotics, a significant proportion had shown resistance against the commonly recommended first-line antibiotics.

scholarly journals Genetic Characterization Among Carbapenem-resistant Escherichia coli Strains Using Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction (ERIC-PCR) Fingerprinting in South Africa

2020 ◽  
Author(s):  
Kingsley Ehi Ebomah ◽  
Anthony Ifeanyi Okoh
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Antimicrobial Agents ◽  
Genetic Relatedness ◽  
Carbapenem Resistance ◽  
Beta Lactam ◽  
Enterobacterial Repetitive Intergenic Consensus ◽  
Chain Reaction ◽  
E Coli ◽  
Polymerase Chain

Abstract Background Carbapenems belong to beta-lactam class of antibiotics usually considered as the last line of defense because they can be effective against severe infections caused by prevalent multidrug-resistant (MDR) pathogens. However, carbapenems can be deactivated by bacteria that produce carbapenemase (beta-lactamase). This study was conducted to screen for carbapenem-resistance genes (CRGs) harbored by pathogenic strains of Escherichia coli recovered from different environmental samples. We also assessed the genetic relatedness among selected E. coli pathotypes using enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR).Method: Molecular identification and characterization of the presumptive isolates were performed using PCR and isolates that exhibited antimicrobial resistance (AMR) phenotypically were further screened for some relevant CRGs (blaNDM−1, blaKPC and blaOXA−48−like). Furthermore, ERIC-PCR was used to determine the similarity and diversity of 31 E. coli strains which were randomly selected from the different sources analyzed in this study.Result Our findings revealed a total of 238 presumptive E. coli isolates, out of which 192 were confirmed positive for uidA gene. Further screening revealed 77 (40%) isolates belong to six key E. coli pathotypes and 70 of them exhibited phenotypic AMR. Additionally, twenty-nine (41%) of the 70 MDR pathogenic E. coli strains harbored CRGs; with 24 strains harboring blaNDM−1, 8 harboring blaKPC and 2 harboring blaOXA−48−like genes.Conclusion Findings also suggest that the selected E. coli pathotypes belonged to different genomic clusters, while the cluster analysis showed a possible genetic diversity among aquatic and farm isolates. Proper treatment of final effluents before discharge as well as the development of more effective strategies to control and manage the use of antimicrobial agents were strongly recommended.

scholarly journals DETEKSI GEN SHV PADA ISOLAT KLINIK Escherichia coli PENGHASIL EXTENDED SPECTRUM BETA−LACTAMASES (ESBLs) DENGAN METODE POLYMERASE CHAIN REACTION (PCR) DARI URIN PASIEN DI RSUD Dr. SOETOMO SURABAYA

2018 ◽  
Vol 11 (2) ◽  
pp. 91-98
Author(s):  
Yulianto Ade Prasetya
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Clinical Microbiology Laboratory ◽  
Beta Lactam ◽  
Chain Reaction ◽  
E Coli ◽  
Beta Lactam Antibiotics ◽  
Extended Spectrum ◽  
Extended Spectrum Beta Lactamases ◽  
Polymerase Chain

 AbstrakEscherichia coli penghasil Extended Spectrum Beta Lactmases (ESBLs) bertanggungjawab terhadap terjadinya wabah infeksi nosokomial, peningkatan morbiditas dan mortalitas, serta peningkatan biaya kesehatan. Enzim yang diproduksi oleh gen SHV dari bakteri mampu menghidrolisis antibiotik sefotaksim dan seftazidim. Tujuan penelitian ini adalah untuk mengetahui prevalensi gen SHV pada isolat klinik E. coli penghasil ESBLs dari urin pasien yang merupakan koleksi Laboratorium Mikrobiologi Klinik RSUD Dr. Soetomo Surabaya pada bulan Januari-Februari 2014. Jenis penelitian yang digunakan adalah observasional deskriptif dengan pendekatan molekuler. Deteksi gen SHV menggunakan metode Polymerase Chain Reaction (PCR) yang kemudian dilakukan elektroforesis dan divisualisasikan pada gel agarose 1,5%. Isolat E. coli yang positif membawa gen SHV ditunjukkan dengan adanya amplikon sebesar 867 bp. Hasil penelitian menunjukkan bahwa dari 30 isolat, sebanyak 12 isolat (40%) positif mengandung gen SHV, dengan prevalensi tertinggi berada di Ruang Instalasi Rawat Jalan. Meropenem dan fosfomisin masih dapat digunakan untuk terapi penyakit yang disebabkan oleh E. coli penghasil ESBLs. Deteksi ESBLs secara genotipik penting dilakukan karena beberapa gen ESBLs menunjukkan resistensi yang berbeda terhadap antibiotik golongan beta laktam. Hasil tersebut memberi informasi kepada pihak rumah sakit manapun untuk mewaspadai prevalensi E. coli penghasil ESBLs melalui pengawasan yang ketat pelaksanaan pemberian antibiotika sesuai tata laksananya.Abstract Extended Spectrum Beta Lactmases (ESBLs) producing−Escherichia coli strains are responsible for the occurrence of nosocomial infection outbreaks, increase of morbidity and mortality, as well as increased healthcare costs. The enzyme produced by the SHV gene from bacteria are able to hydrolyze antibiotics of cefotaxime and cefazazim. The purpose of this study was to detect the presence of SHV gene in clinical ESBLs−producing E. coli isolates from patients’ urines which are collections of Clinical Microbiology Laboratory in Dr. Soetomo Hospital of Surabaya within period of January−February 2014. The research used descriptive observasional design with molecular approach. The SHV gene was detected by using Polymerase Chain Reaction (PCR) method, then the products was performed by electrophoresis and visualized on 1.5% agarose gel. E. coli isolates that positively carrying the SHV gene were demonstrated in the presence of amplicons of 867 bp. The results showed that of 30 isolates, 12 isolates (40%) positively contained the SHV gene, with the highest prevalence being in the Outpatient Installation Room. Meropenem and fosfomycin can still be used for disease therapy caused by ESBLs−producing E. coli. Genotypic detection of ESBLs is important because some ESBLs genes exhibit different resistance to beta−lactam antibiotics. The result provides information to any hospital to be aware of the prevalence of ESBLs−producing E.coli through strict supervision of the implementation of antibiotics according to their administration.

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.

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. 

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.

scholarly journals Prevalence and characteristics ofEscherichia coilserotype O157:H7 and other verotoxin-producingE. coliin healthy cattle

1996 ◽  
Vol 117 (2) ◽  
pp. 251-257 ◽  
Author(s):  
M. Blanco ◽  
J. E. Blanco ◽  
J. Blanco ◽  
E. A. Gonzalez ◽  
A. Mora ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Gene Sequence ◽  
Heterogeneous Population ◽  
Chain Reaction ◽  
E Coli ◽  
Healthy Cattle ◽  
Faecal Samples ◽  
Polymerase Chain

SummaryFrom February to July of 1994, 328 faecal samples from 32 herds were collected and verotoxin-producingEscherichia coli(VTEC) found on 84% of the farms. The proportion of animals infected varied from 0–63%. VTEC were recovered from 52 (20%) of 257 cows and from 16 (23%) of 71 calves. Although the VTEC belonged to 25 different serogroups, 7 (O8. O20, O22, O77, O113, O126 and O162) accounted for 46% of strains. Nearly 45% of the 83 bovine VTEC strains belonged to serogroups associated with haemorrhagic colitis and haemolytic uraernic syndrome in humans. However, only 2 (2%) of 83 VTEC strains isolated from cattle belonged to enterohaemorrhagicE. coli(EHEC) serotypes (O26:H11 and O157:H7), and only 8 (10%) were positive for the attaching and effacingE. coli (eae)gene sequence. Polymerase chain reaction (PCR) showed that 17 (20%) of VTEC strains carried VT1 genes. 43 (52%) possessed VT2 genes, and 23 (28%) carried both VT1 and VT2 genes. Characterization of VTEC isolates revelated a heterogeneous population in terms of serogroup and toxin type in the positive herds. This study confirms that healthy cattle are a reservoir of VTEC, but, the absence ofeaegenes in most bovine VTEC strains suggests that they may be less virulent for humans thaneae-positive EHEC.

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