Detection of Amp C beta lactamases production in Escherichia coli & Klebsiella by an inhibitor based method.

BACKGROUND & OBJECTIVE: 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. METHODS: 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. RESULTS: 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. INTERPRETATION & CONCLUSION: 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.

Detection of metallo betalactamase producing Pseudomonas aeruginosa in hospitalized patients.

BACKGROUND & OBJECTIVE: Metallo beta-lactamase (MBL)-mediated resistance to carbapenems is an emerging threat in hospital isolates of Pseudomonas aeruginosa. Though there are several screening methods to detect this enzyme production, the National Committee for Clinical Laboratory Standards (NCCLS) does not have performance standards documented so far. There is not enough information from the Indian subcontinent regarding the prevalence and the screening methods for these enzymes. The present study was undertaken to detect MBL in nosocomial isolates of P. aeruginosa by two screening methods. METHODS: Fifty consecutive P. aeruginosa isolates obtained from hospitalized patients were subjected to susceptibility testing to antipseudomonal drugs by disc diffusion, and minimum inhibitory concentration (MIC) of imipenem was determined. The production of MBL was detected by 4-fold reduction in MIC with imipenem-ethylene diamine tetraacetic acid (EDTA) and the zone size enhancement with EDTA impregnated imipenem and ceftazidime discs. RESULTS: Sixteen per cent of the isolates tested were resistant to imipenem by disc diffusion method of which 87.5 per cent exhibited a zone size enhancement with EDTA impregnated imipenem and ceftazidime discs as well as a 4-fold reduction in MIC with imipenem EDTA. The imipenem susceptible isolates (84%) had normal MIC values and exhibited no zone diameter enhancement with EDTA impregnated antibiotic discs. INTERPRETATION & CONCLUSION: MBL-mediated imipenem resistance in P. aeruginosa is a cause for concern in the therapy of critically ill patients. The two confirmatory methods i.e., zone diameter enhancement with EDTA impregnated imipenem and ceftazidime discs and 4-fold reduction in MIC with imipenem EDTA combination are equally effective for their detection.

Superantigens - Concepts, clinical disease and therapy.

The term superantigen was introduced in microbiology and immunology only a decade ago. The unique feature of superantigen is that it bypasses the antigen processing mechanism and specifically binds to TCR v segment and forms a trimolecular complex along with major histocompatibility complex class II. Since its discovery, several studies have been carried out to unravel the properties of superantigens. They are implicated in the pathogenesis of diseases of bacterial, viral and fungal origin. They are also known to play a role in autoimmune diseases. Therapy for these superantigens is being worked upon. Studies have shown that intravenous gamma globulin containing specific antibodies to these superantigens inhibits the activation of T-Cells and also the cytokine production by them resulting in dramatic recovery from superantigen mediated diseases. The use of superantigen in the therapy of cancer is being explored.