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 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 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.

scholarly journals Despite Predominance of Uropathogenic/Extraintestinal Pathotypes Among Travel-acquired Extended-spectrum β-Lactamase–producing Escherichia coli, the Most Commonly Associated Clinical Manifestation Is Travelers’ Diarrhea

2019 ◽  
Vol 70 (2) ◽  
pp. 210-218 ◽  
Author(s):  
Anu Kantele ◽  
Tinja Lääveri ◽  
Sointu Mero ◽  
Inka M K Häkkinen ◽  
Juha Kirveskari ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Chain Reaction ◽  
Symptomatic Infection ◽  
E Coli ◽  
Extended Spectrum ◽  
Before And After ◽  
Clinical Consequences ◽  
Polymerase Chain ◽  
Enterobacteriaceae Infections ◽  
The Tropics

AbstractBackgroundOne-third of the 100 million travelers to the tropics annually acquire extended-spectrum β-lactamase (ESBL)–producing Enterobacteriaceae (ESBL-PE), with undefined clinical consequences.MethodsSymptoms suggesting Enterobacteriaceae infections were recorded prospectively among 430 Finnish travelers, 90 (21%) of whom acquired ESBL-PE abroad. ESBL-PE isolates underwent polymerase chain reaction–based detection of diarrheagenic Escherichia coli (DEC) pathotypes (enteroaggregative E. coli [EAEC], enteropathogenic E. coli [EPEC], enterotoxigenic E. coli [ETEC], enteroinvasive E. coli, and Shiga toxin–producing E. coli), and extraintestinal pathogenic/uropathogenic E. coli (ExPEC/UPEC). Laboratory-confirmed ESBL-PE infections were surveyed 5 years before and after travel.ResultsAmong the 90 ESBL-PE carriers, manifestations of Enterobacteriaceae infection included travelers’ diarrhea (TD) (75/90 subjects) and urinary tract infection (UTI) (3/90). The carriers had 96 ESBL-producing E. coli isolates, 51% exhibiting a molecular pathotype: 13 (14%) were DEC (10 EAEC, 2 EPEC, 1 ETEC) (12 associated with TD) and 39 (41%) ExPEC/UPEC (none associated with UTI). Of ESBL-PE, 3 (3%) were ExPEC/UPEC-EAEC hybrids (2 associated with diarrhea, none with UTI). Potential ESBL-PE infections were detected in 15 of 90 subjects (17%). The 10-year medical record survey identified 4 laboratory-confirmed ESBL-PE infections among the 430 travelers, all in subjects who screened ESBL-PE negative after returning home from their index journeys but had traveled abroad before their infection episodes.ConclusionsHalf of all travel-acquired ESBL-producing E. coli strains qualified molecularly as pathogens. Extraintestinal and uropathogenic pathotypes outnumbered enteric pathotypes (41% vs 14%), yet the latter correlated more closely with symptomatic infection (0% vs 92%). Despite more ESBL-PE strains qualifying as ExPEC/UPEC than DEC, travel-acquired ESBL-PE are more often associated with TD than UTI.

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 Characterization of Multidrug Resistant Extended-Spectrum Beta-Lactamase-ProducingEscherichia coliamong Uropathogens of Pediatrics in North of Iran

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Mohammad Sadegh Rezai ◽  
Ebrahim Salehifar ◽  
Alireza Rafiei ◽  
Taimour Langaee ◽  
Mohammadreza Rafati ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Multidrug Resistant ◽  
Beta Lactamase ◽  
Beta Lactam ◽  
Extended Spectrum Beta Lactamase ◽  
E Coli ◽  
Beta Lactam Antibiotics ◽  
Extended Spectrum ◽  
Extended Spectrum Beta Lactamases ◽  
North Of Iran

Escherichia coliremains as one of the most important bacteria causing infections in pediatrics and producing extended-spectrum beta-lactamases (ESBLs) making them resistant to beta-lactam antibiotics. In this study we aimed to genotype ESBL-producingE. coliisolates from pediatric patients for ESBL genes and determine their association with antimicrobial resistance. One hundred of theE. coliisolates were initially considered ESBL producing based on their MIC results. These isolates were then tested by polymerase chain reaction (PCR) for the presence or absence ofCTX,TEM,SHV,GES, andVEBbeta-lactamase genes. About 30.5% of isolatedE. coliwas ESBL-producing strain. TheTEMgene was the most prevalent (49%) followed bySHV(44%),CTX(28%),VEB(8%), andGES(0%) genes. The ESBL-producingE. coliisolates were susceptible to carbapenems (66%) and amikacin (58%) and showed high resistance to cefixime (99%), colistin (82%), and ciprofloxacin (76%). In conclusion, carbapenems were the most effective antibiotics against ESBl-producingE. coliin urinary tract infection in North of Iran. The most prevalent gene is the TEM-type, but the other resistant genes and their antimicrobial resistance are on the rise.

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