scholarly journals Molecular Characterization of blaCTX-M, blaTEM and blaSHV Beta Lactamases Produced by Uropathogens

2020 ◽  
pp. 97-106
Author(s):  
Ashna Bhasin ◽  
Yogesh Chander ◽  
Harmesh Manocha
Keyword(s):  
Polymerase Chain Reaction ◽  
Test Method ◽  
Control Measures ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Esbl Production ◽  
Chain Reaction ◽  
Disk Diffusion Test ◽  
Polymerase Chain ◽  
Tract Infections

Background: This prospective study was carried out to look for the frequency of isolation of Extended spectrum b lactamase (ESBL) producing bacteria from urine samples and study their susceptibility pattern. The detection of ESBL genes responsible for their resistance was done by Polymerase chain reaction (PCR). Methods: The study was carried out over a period of one year from January 2016 to        December 2016. Urine specimens from patients were processed as per standard protocol. Antimicrobial susceptibility testing was performed by disk diffusion method as per CLSI guidelines 2016. Urine isolates obtained were screened for ESBL, by cefotaxime, ceftazidime disk and confirmation was made by Double disk diffusion test method. The detection of ESBL genes responsible for their resistance was done by Polymerase chain reaction (PCR) for blaCTX-M, blaTEM and blaSHV genes. Results: Prevalence of ESBL producing uropathogens were found to be 20.47% with most common organism to be isolated was Escherichia coli (E. coli) followed by Klebsiella pneumoniae. Nitrofurantoin and Imipenem were the most effective antibiotic agents against urinary isolates. Most common gene responsible for ESBL production was blaCTX-M (71.42%). Conclusion: A large number of ESBL producing strains are creating significant therapeutic problems. Therefore, monitoring of ESBL production, judicious use of antibiotics and infection control measures are necessary to avoid treatment failures in patients with Urinary Tract Infections (UTI).

scholarly journals Identification of VIM and IMP genes and metallo-betalactamase enzymes in Escherichia coli isolates by molecular and phenotypic methods in shahrekord educational hospitals

2020 ◽  
Vol 22 (1) ◽  
pp. 46-52
Author(s):  
Farshad Kakian ◽  
Kourosh Naderi ◽  
Mohamad Hosaein Rezaei ◽  
Majid Validi ◽  
Behnam Zamanzad ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Multiple Drug Resistance ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Chain Reaction ◽  
E Coli ◽  
Polymerase Chain ◽  
Tract Infections ◽  
Phenotypic Tests

Background and aims:: Among urine pathogens, Escherichia coli (E. coli) causes 80% of urinary tract infections (UTIs). Due to the destructive nature of penicillins, cephalosporins and carbapenems (except for monobactam such as aztreonam) and carbapenemase enzymes have created many problems for treating infectious diseases. Therefore, this study aimed to investigate the phenotypic and molecular characterization of metallo-beta-lactamase (MBL) genes produced by E. coli isolates in an educational hospital during 2016- 2017. Methods: This cross-sectional study investigated 80 UTI samples affected by E. coli. In addition, antibiotic susceptibility was evaluated by disk diffusion and E-test methods for two antibiotics of meropenem and imipenem. Phenotypic tests containing modified Hodge test, ethylenediaminetetraacetic acid (EDTA) disk synergy test, and AmpC Disk were performed to identify MBL enzyme-producing strains. Finally, the frequency of Verona integron-encoded metallo-β-lactamase (VIM) and imipenemase (IMP) genes was determined by polymerase chain reaction (PCR). Results: Among 80 E. coli samples, 21 (26.25%) isolates were resistant to meropenem and imipenem as detected by the disk-diffusion method and E-test. Further, phenotypic tests including modified Hodge test, EDS test, and AmpC disk test showed the positivity of 15 (18.75%), 15 (18.75%), and 8 (10%) isolates, respectively (P < 0.001). Eventually, polymerase chain reaction (PCR) test results for the VIM gene showed 19 (23.75%) positive isolates of E. coli, but the IMP gene was observed in none of the isolates (P<0.001). Conclusion: In general, the emergence of E. coli producing MBL enzymes is a serious threat among clinical infections. The findings of this study indicated the presence of E. coli producing MBL. These enzymes can degrade carbapenems antibiotics, the last class current treatment of multiple drug-resistance infections.

Urinary Tract Infections in Patients with Type 1 and Type 2 Diabetes: Etiology, Resistance to Antibacterial Chemicals and Virulence Features

2017 ◽  
Vol 68 (3) ◽  
pp. 566-569 ◽  
Author(s):  
Mihaela Magdalena Mitache ◽  
Carmen Curutiu ◽  
Elena Rusu ◽  
Ramona Bahna ◽  
Mara Ditu ◽  
...  
Keyword(s):  
Urinary Tract ◽  
Virulence Factors ◽  
Urinary Tract Infections ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Diabetic Patients ◽  
Disk Diffusion Test ◽  
Tract Infections ◽  
Susceptibility Profiles ◽  
Diabetes Patients

One of the most frequent chronic complications occurred in diabetes patients are the urinary tract infections (UTIs). This study aimed to investigate the incidence of UTIs in a cohort of 93 (47 males: 46 females) diabetic patients, the prevalence of different microbial species involved and their virulence and antibiotic resistance profiles. The identification of the uropathogenic strains in the positive urine samples was performed using conventional methods and API tests. After identification, the antibiotic susceptibility profiles were established by the standardized disk diffusion method and double disk diffusion test was performed for the confirmation of ESBL and inducible AmpC b �lactamase phenotypes. The isolated strains were tested for the production of different cell associated and soluble virulence factors, i.e.: bacterial adherence to cellular substrata (HeLa cells), hemolysins (hemolysis spot, CAMP-like), amylase, caseinase, aesculin hydrolysis, DNA-ase, lipase and lecithinase. In the analyzed group, the total prevalence of UTIs was of 46%, a higher incidence being observed in the female patients (64%). Similar to other studies, the etiology of UTI in the investigated diabetes patients was dominated by E. coli, followed by Klebsiella sp. strains. The isolated strains preserved good susceptibility rates to quinolones and aminoglycosides and revealed important virulence features, related to their capacity to colonize the cellular substratum and to produce soluble virulence factors involved in persistence, colonization and progression of the infectious process. The high percentage of beta-lactam resistant strains (including carbapenem-resistant ones) requires careful surveillance of the dynamics of susceptibility profiles for limiting the emergence of these strains in community.

Molecular Analysis of Uropathogenic E.coli Isolates from Urinary Tract Infections

2019 ◽  
Vol 19 (3) ◽  
pp. 322-326 ◽  
Author(s):  
Hassan Valadbeigi ◽  
Elham Esmaeeli ◽  
Sobhan Ghafourian ◽  
Abbas Maleki ◽  
Nourkhoda Sadeghifard
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Urinary Tract ◽  
Urinary Tract Infections ◽  
Multiplex Polymerase Chain Reaction ◽  
Virulence Genes ◽  
Uropathogenic Escherichia Coli ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Tract Infections

Introduction: The aim of the current study was to investigate the prevalence of virulence genes in uropathogenic Escherichia coli (UPEC) isolates in Ilam. Materials and Methods: For this purpose, a total of 80 UPEC isolates were collected for patients with UTIs during a 6 months period. The multiplex polymerase chain reaction (multiplex PCR) was used to detect the papEF, fimH, iucD, hlyA, fyuA, and ompT genes. Results: The prevalence of fimH, papEF, iucD, fyuA, hlyA, hlyA, and ompT genes were 87.5%, 47.5%, 60%, 67.5%, 27.5%, 47.5% and 71.2%, respectively. Among all of the isolates, 27 profiles were obtained. Conclusion: Our findings demonstrated that the most prevalence was found for fimH, and different distribution of virulence genes suggested different ability of pathogenicity.

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 USE OF THE POLYMERASE CHAIN REACTION FOR THE DIAGNOSIS OF ASYMPTOMATIC Leishmania INFECTION IN A VISCERAL LEISHMANIASIS-ENDEMIC AREA

2013 ◽  
Vol 55 (2) ◽  
pp. 101-104 ◽  
Author(s):  
Luciana Almeida Silva ◽  
Héctor Dardo Romero ◽  
Aline Fagundes ◽  
Nédia Nehme ◽  
Otávio Fernandes ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Visceral Leishmaniasis ◽  
Endemic Area ◽  
Asymptomatic Infection ◽  
Control Measures ◽  
Skin Tests ◽  
Leishmania Infection ◽  
Chain Reaction ◽  
Serological Tests ◽  
Polymerase Chain

The diagnosis of asymptomatic infection with Leishmania (Leishmania) chagasi has become more important over recent years. Expansion of visceral leishmaniasis might be associated with other routes of transmission such as transfusion, congenital or even vector transmission, and subjects with asymptomatic infection are potential reservoirs. Moreover, the identification of infection may contribute to the management of patients with immunosuppressive conditions (HIV, transplants, use of immunomodulators) and to the assessment of the effectiveness of control measures. In this study, 149 subjects living in a visceral leishmaniasis endemic area were evaluated clinically and submitted to genus-specific polymerase chain reaction (PCR), serological testing, and the Montenegro skin test. Forty-nine (32.9%) of the subjects had a positive PCR result and none of them developed the disease within a follow-up period of three years. No association was observed between the results of PCR, serological and skin tests. A positive PCR result in subjects from the endemic area did not indicate a risk of progression to visceral leishmaniasis and was not associated with a positive result in the serological tests.

scholarly journals Prescription Surveillance and Polymerase Chain Reaction Testing to Identify Pathogens during Outbreaks of Infection

2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Hiroaki Sugiura ◽  
Tsuguto Fujimoto ◽  
Tamie Sugawara ◽  
Nozomu Hanaoka ◽  
Masami Konagaya ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Syndromic Surveillance ◽  
Respiratory Infections ◽  
Emerging Infectious Diseases ◽  
Upper Respiratory Tract ◽  
Chain Reaction ◽  
Specific Pathogen ◽  
Polymerase Chain ◽  
Syncytial Virus ◽  
Tract Infections

Syndromic surveillance, including prescription surveillance, offers a rapid method for the early detection of agents of bioterrorism and emerging infectious diseases. However, it has the disadvantage of not considering definitive diagnoses. Here, we attempted to definitively diagnose pathogens using polymerase chain reaction (PCR) immediately after the prescription surveillance system detected an outbreak. Specimens were collected from 50 patients with respiratory infections. PCR was used to identify the pathogens, which included 14 types of common respiratory viruses andMycoplasma pneumoniae. Infectious agents includingM. pneumoniae, respiratory syncytial virus (RSV), rhinovirus, enterovirus, and parainfluenza virus were detected in 54% of patients. For the rapid RSV diagnosis kit, sensitivity was 80% and specificity was 85%. For the rapid adenovirus diagnosis kit, no positive results were obtained; therefore, sensitivity could not be calculated and specificity was 100%. Many patients were found to be treated for upper respiratory tract infections without the diagnosis of a specific pathogen. In Japan, an outbreak ofM. pneumoniaeinfection began in 2011, and our results suggested that this outbreak may have included false-positive cases. By combining syndromic surveillance and PCR, we were able to rapidly and accurately identify causative pathogens during a recent respiratory infection outbreak.

Accuracy of real-time polymerase chain reaction to detect Schistosoma mansoni – infected individuals from an endemic area with low parasite loads

Parasitology ◽  
2020 ◽  
Vol 147 (10) ◽  
pp. 1140-1148
Author(s):  
Fernanda do Carmo Magalhães ◽  
Samira Diniz Resende ◽  
Carolina Senra ◽  
Carlos Graeff-Teixeira ◽  
Martin Johannes Enk ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Schistosoma Mansoni ◽  
Real Time ◽  
Diagnostic Methods ◽  
Control Measures ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Cross Sectional ◽  
Intestinal Schistosomiasis ◽  
Polymerase Chain

AbstractDue to the efforts to control schistosomiasis transmission in tropical countries, a large proportion of individuals from endemic areas present low parasite loads, which hinders diagnosis of intestinal schistosomiasis by the Kato-Katz (KK) method. Therefore, the development of more sensitive diagnostic methods is essential for efficient control measures. The aim was to evaluate the accuracy of a real-time polymerase chain reaction (RT-PCR) to detect Schistosoma mansoni DNA in fecal samples of individuals with low parasite loads. A cross-sectional population-based study was conducted in a rural community (n = 257) in Brazil. POC-CCA® was performed in urine and feces were used for RT-PCR. In addition, fecal exams were completed by 18 KK slides, saline gradient and Helmintex techniques. The combined results of the three parasitological tests detected schistosome eggs in 118 participants (45.9%) and composed the consolidated reference standard (CRS). By RT-PCR, 117 out of 215 tested samples were positive, showing 91.4% sensitivity, 80.2% specificity and good concordance with the CRS (kappa = 0.71). RT-PCR identified 86.9% of the individuals eliminating less than 12 eggs/g of feces, demonstrating much better performance than POC-CCA® (50.8%). Our results showed that RT-PCR is a valuable alternative for the diagnosis of intestinal schistosomiasis in individuals with very low parasite loads.

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