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Chaudhary M.,Venus Medicine Research Center | Payasi A.,Venus Medicine Research Center
Journal of Proteomics and Bioinformatics | Year: 2012

The aim of the present investigation was to characterize the prevalence of extended-spectrum β-lactamases (ESBLs) and metallo β-lactamases (MBLs), and to study the antibiotic susceptibility profile among 250 clinical isolates of Acinetobacter baumannii. Phenotypic characterization was carried out by double disc synergy method and the prevalence of ESBLs and MBLs antibiotic resistant determinants were analyzed with Polymerase Chain Reaction (PCR). Susceptibility studies were performed by disc diffusion method according to Clinical and Laboratory Standards Institute guidelines 2009. Among the two hundred fifty isolates, two hundred nine isolates (83.6%) were positive for ESBLs whereas one hundred sixty seven isolates (79.9%) were positive for both ESBLs and MBLs. Moreover, five isolates (2.3%) which were positive for MBL on disc diffusion test, but negative in PCR showed MBL activity by spectrophotometric assay. Susceptibility study showed that all of the isolates were found to be more susceptible to ceftriaxone plus ethylenediaminetetraacetate plus sulbactam (90-93%), followed by meropenem (50-53%), imipenem (42-45%), cefoperazone plus sulbactam (40-42%), piperacillin plus tazobactam (38-42%) and amoxicillin plus clavulanic acid (28-31%). Among the ESBLs, TEM-types were varied from 82 to 87% followed by SHV-types (67-78%), CTX-M types (60 to 67) and OXA types (51 to 56%) in all of the isolates. Among the MBLs, NDM-1 varied from 40 to 49% followed by IMP-1 (51 to 55%), VIM-1 (55 to 59%) and KPC (47 to 55%) in all of the isolates. Moreover, results of the present study revealed that all of the clinical isolates were susceptible to ceftriaxone plus EDTA plus sulbactam and can be a potent antibacterial agent for the treatment of severe bacterial infections caused by A. baumannii. © 2012 Chaudhary M, et al. Source


Chaudhary M.,Venus Medicine Research Center | Payasi A.,Venus Medicine Research Center
American Journal of Infectious Diseases | Year: 2014

In the present study, anaerobic clinical isolates of Bacteroides fragilis, Escherichia coli, Staphylococcus aureus and Yersinia enterocolitica were obtained from different clinical specimens and were subjected to molecular typing to detect the gene encoding metronidazole resistance in these isolates. Subsequently, antibacterial activity of drugs was tested against the selected clinical isolates. A total of 53 clinical isolates involving 18 obligate and 35 facultative anaerobic bacteria were recovered from clinical samples of 67 patients who were suspected to have anaerobic infection. A disk diffusion method was employed to screen for metronidazole-resistance among these isolates. PCR assay was used to detect the metronidazole resistant gene (nim). Susceptibility studies in metronidazole resistant clinical isolates as well as positive controls were performed according to Clinical and Laboratory Standards Institute (CLSI) guidelines. According to disc diffusion method, of 53 isolates, 21 isolates (39.6%) were found to be metronidazole resistant. Further screening of these isolates with PCR revealed only 13 isolates (24.5%) carry nim gene. Out of which 7 were of B. fragilis, 3 were of Y. enterocolitica, 2 were of E. coli and 1 was of S. aureus. The highest number of metronidazole resistant isolates were found in abscess (7) followed by intra-abdominal infection (5) and bone and joint infection (1). When metronidazole resistant isolates were subjected to screen for the presence of nim gene, all isolates were found to carry nim gene. According to minimum inhibitory concentration (MIC) data, among the tested antibacterial agents, Mebatic emerged as the most active antibacterial against metronidazole resistant isolates of B. fragilis, E. coli, S. aureus and Y. enterocolitica with MIC values 0.125 to 1.0 μg mL-1. Similarly, Antimicrobial Susceptibility Test (AST) data also revealed that Mebatic was most efficacious in the metronidazole resistant organisms. From the above results, it is evident that Mebatic has enhanced in vitro antibacterial activity compared to other drugs in metronidazole resistant isolates thus can be a potent antibacterial agent for the treatment of infections caused by metronidazole resistant organisms. © 2014 Science Publication. Source


Chaudhary M.,Venus Medicine Research Center | Payasi A.,Venus Medicine Research Center
Journal of Proteomics and Bioinformatics | Year: 2013

The rising antibiotic resistance against commonly used drugs is of great concern. Drug susceptibility testing and Polymerase Chain Reaction (PCR) assay were used to determine the antibiotic susceptibility patterns and prevalence of genes encoding extended-spectrum-β-lactamases (ESBLs) and metallo-β-lactamases (MBLs) among 515 isolates of Pseudomonas aeruginosa isolated from various clinical specimens. Susceptibility of isolates to seven antibiotics was tested using disc diffusion method according to the guidelines defined by Clinical Laboratory Standard Institute. Isolates showing resistance to any of the two cephalosporins (ceftriaxone, ceftazidime and cefotaxime) were subjected to PCR for the prevalence of ESBL and MBL gene characterization. Out of the 515 isolates, 235 (45.63%) were considered as ESBL positive; 87 (16.89%) were MBL positive and 74 (14.36%) had co-produced both ESBL and MBL. The frequency of TEM-type, SHV-type and AMP-C type ESBLs were 45.10, 26.0, and 28.93%, respectively. Among the MBLs, the frequency of distribution of NDM-1, IMP-1 and VIM-1 was 24.13, 28.73 and 47.12%, respectively. The rate of susceptibility of ESBL producing P. aeruginosa towards various antibacterial agents were as follows: piperacillin+tazobactam (84.3%), doripenem (83.8%), ceftriaxone plus ethylenediamminetetraacetic acid plus sulbactam; Elores (74.1%), imipenem (66.5%), meropenem (54.7%), ceftazidime (44.8%) and cefepime (28.5%). Isolates harboring MBL and ESBL+MBL genes were resistant to almost all antibiotics except Elores (97.3 and 95.1% susceptibility) and doripenem (11.3 and 19.5% susceptibility). From the above results, it can be concluded that Elores was highly potent against MBL producing P. aeruginosa. However, suceptibility of Elores to ESBL producing P. aeruginosa was comparable to piperacillin plus tazobactam. © 2013 Chaudhary M, et al. Source


Chaudhary M.,Venus Medicine Research Center | Payasi A.,Venus Medicine Research Center
American Journal of Infectious Diseases | Year: 2013

Multidrug resistant Methicillin-Resistant Staphylococcus aureus (MRSA) is a major cause of nosocomial and community acquired infections and is on the rise. The aim of this investigation was to explore the prevalence of MRSA and heterogeneous Glycopeptide Intermediate Staphylococcus aureus (hGISA) in various clinical samples, to investigate the various antibiotic resistant determinant genes among these strains collected from north and west Indian hospitals and to evaluate the response of various drugs to these strains. A total of 413 clinical specimens collected from different hospitals were processed for the screening of S. aureus and MRSA. All the MRSA strains were further screened for hGISA on Mueller-Hinton agar containing 8 μg mL-1 teicoplanin or 6 μg mL-1 vancomycin. hGISA confirmed by the E-test method with a dense inoculum and a simplified method of population analysis. Susceptibility study was conducted according to the Clinical and Laboratory Standards Institute (CLSI) methods. Among 211/413 S. aureus clinical isolates, 61.6% (130/211) of the isolates were confirmed to be MRSA which included maximum isolates from pus, blood, urine, wound swab and ear swab samples in decreasing order. hGISA strains were found in 8/130 (6.1%) isolates. Vancoplus, a novel antibiotic adjuvant entity was found to be susceptible in 96.1 to 97.8% MRSA strains and showed intermediate response in 2.2 to 3.8% of isolates. Linezolid appeared to be second most active antibiotic with 48.0 to 81.2% susceptibility, followed by teicoplanin (41.3 to 56.2% susceptibility). There was 8.7 to 9.6% resistance observed in Linezolid which was increased to 48% in teicoplanin, to >60% in daptomycin and >75% in vancomycin. Interestingly, none of the isolates were susceptible to ceftriaxone and cefoperazone plus sulbactam. From the above study it can be concluded that prevelance of MRSA has reached a significant level and Vancoplus is the most effective drug in MRSA as well as hGISA organisms in comparison to comparator drugs. © 2013 Science Publication. Source


Chaudhary M.,Venus Medicine Research Center | Payasi A.,Venus Medicine Research Center
Journal of Microbial and Biochemical Technology | Year: 2012

The aim of the present work was to study the prevalence of Extended-spectrum β-lactamases (ESBLs) and Metallo-β-lactamases (MβLs) among 464 E. coli clinical isolates obtained from various clinical specimens; and to study the susceptibility of various drugs against E. coli isolates. Phenotypic characterization and susceptibility studies were performed according to the methods described in Clinical and Laboratory Standards Institute guidelines (CLSI, 2010). The prevalence of ESBLs and MβLs was analyzed with Polymerase Chain Reaction (PCR), using the previously reported primers. Among the four hundred sixty four isolates, 186 (40.08%) isolates were ESBLs positive, 75 (16.16%) isolates were MβLs positive, and 80 (17.24%) were both ESBLs and MβLs positive. The remaining 123 (26.50%) were non ESBLs and MβLs. TEM-types ESBLs (blaTEM-1, blaTEM-2, and blaTEM-50) were found in approximately 57% isolates. The prevalence of SHV-types, CTX-M-types and OXA-type was 29.03, 11.82 and 2.15%, respectively. Among the MβLs, the frequency of distribution of NDM-1, IMP-1, VIM-1 and KPC-types was 37.39, 21.33, 18.66, and 22.66%, respectively. In general, 92.6% E. coli isolates were susceptible to ceftriaxone plus EDTA plus sulbactam (CSE1034), followed by meropenem (74.4%), imipenem (71.2%), piperacillin plus tazobactam (52.1%), cefoperazone plus sulbactam (46.0%) and amoxycillin plus clavulanic acid (23.6%). Similarly, amoxycillin plus clavulanic acid showed the highest percentage of resistance (72.8%), followed by cefoperazone plus sulbactam (43.6%), piperacillin plus tazobactam (39.3%), imipenem (23.3%), meropenem (20.3%) and ceftriaxone plus EDTA plus sulbactam (CSE1034) (2.5%). Results of the present study revealed that most of the clinical isolates were susceptible to ceftriaxone plus EDTA plus sulbactam (CSE1034), and can be a potent antibacterial agent for the treatment of severe bacterial infections caused by E. coli.© 2012 Manu C, et al. Source

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