Molecular epidemiology of clinical isolates of ampc producing Klebsiella pneumoniae.
ABSTRACT AmpC producing K. pneumoniae have been increasingly reported from India but epidemiological studies are lacking. In the present study, molecular epidemiology of extended-spectrum AmpC beta-lactamases (ESACs) producing clinical isolates of K. pneumoniae prevalent in our hospital was studied.
Fifty-one non-repeat, consecutive, clinical isolates of K. pneumoniae producing AmpC enzymes, were subjected to whole cell protein profile analysis (SDS-PAGE) and ribotyping. The antimicrobial susceptibility was determined using standard disk diffusion technique. The isolates showing decreased susceptibility to cefoxitin (< 18 mm) or cefotetan (< 16 mm) were subjected to modified three- dimensional test for detection of AmpC enzyme.
Six different types of protein profiles were observed. Ribotyping could further discriminate between the strains that were clustered by protein fingerprinting. Twelve different ribo-patterns were identified. Ribotyping was found to have a better Discriminatory Index (0.98) than that of SDS-PAGE (0.78). Of the 26 isolates that showed decreased susceptibility to cefoxitin and/or cefotetan 13 isolates were found to harbour AmpC enzyme.
The study demonstrated the usefulness of SDS-PAGE whole cell protein profile analysis and ribotyping to identify the clonality of the ESACs isolates, the latter having a higher discriminatory power. The presence of ESACs isolates in the community as well as in hospital settings emphasizes the need for regular monitoring of antimicrobial resistance.
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ABSTRACT: Detection of AmpC-mediated resistance in Gram-negative organisms poses a problem due to misleading results in phenotypic tests. There are no recommended guidelines for detection of this resistance mechanism and there is a need to address this issue as much as the detection of extended spectrum beta lactamases (ESBLs) since both may co-exist and mask each other. Though resistance to cefoxitin is used as a screening test, it does not reliably indicate Amp C production. This study was undertaken to detect Amp C beta lactamases in certain Gram-negative bacteria employing an inhibitor base test using boronic acid. A total number of 76 consecutive non repetitive clinical isolates of Escherichia coli (n=67) and 9 Klebsiella pneumoniae (n=9) obtained over a period of two months, were screened for amp C production by disc diffusion method using cefoxitin (30 microg) dics and confirmed by inhibitor based test using boronic acid as inhibitor. A total of 36 of 76 isolates (47.3%) screened harboured amp C enzymes, of which a majority 31 (86.1%) co-produced ESBL enzymes. Pure ampC production was seen in 7 (9.2%) of isolates only. Most of the amp C producers also produced ESBL enzymes. The inhibitor based test was useful in identifying cefoxitin susceptible amp C producers and could also effectively differenciate ESBL from amp C producing isolates.The Indian Journal of Medical Research 09/2007; 126(3):220-3. · 2.06 Impact Factor
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ABSTRACT: AmpC β-lactamases are cephalosporinases that hydrolyze cephamycins as well as other extended-spectrum cephalosporins and are poorly inhibited by clavulanic acid. Although reported with increasing frequency, the true rate of occurrence of AmpC β-lactamases in different organisms, including members of Enterobacteriaceae, remains unknown. The present study was designed to determine the occurrence of AmpC enzyme-harbouring Gram-negative clinical isolates in a tertiary care hospital in Pondicherry state, South India. A total of 235 Gram negative clinical isolates were tested for resistance to cefoxitin, third generation cephalosporin (3GC) antibiotics, ampicillin, amikacin, co-trimoxazole, gentamicin, meropenem and tetracycline by disc diffusion method. Isolates found resistant to 3GC and cefoxitin were tested for the production of AmpC β -lactamases by three dimensional extraction method and AmpC disc method. Isolates found to sensitive to 3GC were subjected to disc antagonism test for inducible AmpC production. One hundred and thirty four (57%) strains were resistant to 3GC, among which 63(47%) were positive for plasmid-mediated AmpC beta lactamases production. Among the 101 strains sensitive to 3GC, 23 (22.7%) revealed the presence of inducible AmpC beta lactamases by disc approximation test. A total of 80.9% (51/63) of screen positive isolates were detected by Amp C disc test and 93.6% (59/63) by three dimensional extraction method. Out of the 86 AmpC producers, 67 (77.9%) were cefoxitin resistant .Inducible AmpC was not found in Esch.coli and Klebsiella spp. The AmpC producers also concurrently showed multidrug resistance pattern. AmpC producers were found to be prevalent in our hospital and though three dimensional extraction test detects AmpC better, the disk test is easier to perform routinely and is user- friendly.Brazilian Journal of Microbiology 07/2010; 41(3):596-602. · 0.76 Impact Factor
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ABSTRACT: Resistance to higher antimicrobial agent is commonly seen in gram negative bacilli. This issue is a challenging problem to the medical practitioners in addition to it is financial impact on the health care system. To document the prevalence of multi drug resistant gram negative bacilli isolated from urine of patients attending the Urology Department of Tertiary care Hospital of western India in year 2008. Out of total 328 isolates, 118 (35.98%) E.coli, 72 (21.95 %) Klebsiella, 64 (19.51%) Pseudomonas aeruginosa, 30 (9.15%) Acinetobacter, 18 (5.49%) Proteus vulgaris, 18 (5.49%) Proteus mirabilis, 6 (1.83%) Providencia rettgerii, 2 (0.61%) Citrobacter freundii. Out of these isolates, 228 (69.51%) were β-lactamase positive, while 100 (30.51%) were β-lactamase negative. Out of 228 β-lactamase positive, 104 (45.61%) were AmpC β-lactamase positive. Stringent protocol such as Antibiotic policy and Hospital infection control program are mandatory to curb these microbes in a tertiary care hospital.Urology Annals 01/2010; 2(1):7-11.