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2.
Indian J Med Microbiol ; 2015 Jan-Mar ; 33 (1): 21-24
مقالة ي الانجليزية | IMSEAR | ID: sea-156984

الملخص

Purpose: Linezolid is an effective drug against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE). We describe the emergence of linezolid resistance in MRSA and VRE from India. Material and Methods: One MRSA and two VRE strains were isolated from a patient on linezolid therapy of one week duration. All three isolates were resistant to linezolid with minimal inhibitory concentrations (MIC) ≥4 mg/L. The 746-bp region fl anking the possible G2576U mutation on the corresponding DNA from the 23S rRNA was amplifi ed by polymerase chain reaction (PCR) and amplicons were sequenced for all the three isolates. Conjugation experiments using the linezolid resistant MRSA (LRMRSA) and linezolid resistant VRE (LRVRE) isolates as donors and wild strains of corresponding genera as recipients were performed. Results: The MRSA isolate had the classical G2576U mutation. High quality value scores in the sequencing software validated the mutation. Conjugation studies did not indicate presence of transferable resistance for linezolid. Sequencing did not indicate presence of any mutation in the two LRVRE isolates. Conclusions: This is the fi rst report from India citing resistance in Staphylococcus and Enterococcus against Linezolid.

3.
Indian J Med Microbiol ; 2013 Jul-Sept; 31(3): 237-241
مقالة ي الانجليزية | IMSEAR | ID: sea-148090

الملخص

Purpose: Carbapenem resistance in Acinetobacter baumannii has become highly rampant, which has been ascribed to the presence of multiple carbapenemases. The objective of the present study was to prospectively investigate the presence of multiple carbapenemase encoding genes in clinical isolates of A. baumannii. Materials and Methods: A total of 30 imipenem resistant, consecutive non-repeat clinical isolates A. baumannii from a Tertiary Care Centre of Delhi were subjected to antimicrobial susceptibility testing (AST), screening for carbapenemase production by modified Hodge test (MHT) and determination of minimum inhibitory concentration for imipenem by E-Test® . These were subjected to Real time PCR for blaIMP-1 and 2 , blaVIM-1 and 2 , blaOXA23, 24, 51 and 58 using SYBR green-I. These were grouped together on the basis of their genotype as each isolate harboured multiple carbapenemases and correlated with their AST profile. Detection of the novel carbapenemase blaNDM-1 was performed by real time PCR using TaqMan probes on 14 isolates. Results: Colistin appeared to be the most effective drug in vitro, followed by tetracycline and beta lactam/beta lactamase inhibitor combinations. All, but one isolate were positive for the MHT. All 30 isolates were positive for blaOXA-51 like gene as well as blaIMP-1 and blaVIM-1 genes. blaOXA 24 and 58 were not detected in any of the isolates. blaIMP-2 , blaVIM-2 , blaOXA-23 were present in 15, 6 and 14 isolates respectively. Grouping based on the genotypic profile did not correlate with susceptibility pattern. Nine among the 14 isolates also harboured the novel blaNDM-1 gene. Conclusions: This is the first study from North India, which comprehensively detected the presence of multiple carbapenemases as well the blaNDM-1 gene. The presence of the novel gene blaNDM-1 indicated ability of A. baumannii to acquire new carbapenemase genes despite the existence of multiple carbapenemase genes. The present study confirmed the presence of multiple genetic mechanisms for carbapenemases production among the clinical isolates of A. baumannii in north India.

4.
Indian J Med Microbiol ; 2011 Jul-Sept; 29(3): 275-279
مقالة ي الانجليزية | IMSEAR | ID: sea-143833

الملخص

Purpose: The emergence and spread of zinc-dependent carbapenem resistance has become a diagnostic challenge for clinical microbiologists. The objective of the present study was to screen zinc-dependent carbapenemase activity in clinical isolates of family Enterobacteriaceae. Materials and Methods: A total of 102 multidrug-resistant organisms (MDROs), non-repeat clinical isolates of family Enterobacteriaceae from two tertiary care centres in Delhi, were screened for carbapenemase production by a modified Hodge test (MHT) and additionally by a re-modified Hodge test, EDTA double disc synergy test, and combined disc test (or disc enhancement test) to determine zinc dependence of carbapenemases harbouring bacteria. Results: Of the total 102 clinical isolates (June through November 2010), 91 were from urine and 11 were from blood specimens. The isolates were obtained from patients visiting the outpatient department (18 isolates), admitted in non-ICU inpatient care units (74 isolates) and patients admitted in ICUs (4 isolates). MHT identified 92 (90.2%) isolates as carbapenemases producers. Among those found negative for MHT (n=10), metallo-beta-lactamases (MBLs) activity was demonstrated through the EDTA disc diffusion synergy test and the combined disc test in 8 and 9 isolates respectively. A total of 63 (61.7%) isolates demonstrated MBL activity despite in vitro sensitivity to Imipenem. Conclusions: The study demonstrated that supplementing the MHT with at least one of the screening methods increases the likelihood of picking up such isolates that may be missed by the MHT. The study also demonstrates the wide-spread presence of MBLs in Enterobacteriaceae members from patients visiting hospitals in east Delhi.


الموضوعات
Bacterial Proteins/metabolism , Bacteriological Techniques/methods , Coenzymes/metabolism , Drug Resistance, Multiple, Bacterial , Enterobacteriaceae/enzymology , Enterobacteriaceae/isolation & purification , Enterobacteriaceae Infections/microbiology , Hospitals , Humans , Immunoassay/methods , India , Zinc/metabolism , beta-Lactamases/metabolism
5.
Indian J Med Microbiol ; 2011 Jul-Sept; 29(3): 275-279
مقالة ي الانجليزية | IMSEAR | ID: sea-143830

الملخص

Purpose: The emergence and spread of zinc-dependent carbapenem resistance has become a diagnostic challenge for clinical microbiologists. The objective of the present study was to screen zinc-dependent carbapenemase activity in clinical isolates of family Enterobacteriaceae. Materials and Methods: A total of 102 multidrug-resistant organisms (MDROs), non-repeat clinical isolates of family Enterobacteriaceae from two tertiary care centres in Delhi, were screened for carbapenemase production by a modified Hodge test (MHT) and additionally by a re-modified Hodge test, EDTA double disc synergy test, and combined disc test (or disc enhancement test) to determine zinc dependence of carbapenemases harbouring bacteria. Results: Of the total 102 clinical isolates (June through November 2010), 91 were from urine and 11 were from blood specimens. The isolates were obtained from patients visiting the outpatient department (18 isolates), admitted in non-ICU inpatient care units (74 isolates) and patients admitted in ICUs (4 isolates). MHT identified 92 (90.2%) isolates as carbapenemases producers. Among those found negative for MHT (n=10), metallo-beta-lactamases (MBLs) activity was demonstrated through the EDTA disc diffusion synergy test and the combined disc test in 8 and 9 isolates respectively. A total of 63 (61.7%) isolates demonstrated MBL activity despite in vitro sensitivity to Imipenem. Conclusions: The study demonstrated that supplementing the MHT with at least one of the screening methods increases the likelihood of picking up such isolates that may be missed by the MHT. The study also demonstrates the wide-spread presence of MBLs in Enterobacteriaceae members from patients visiting hospitals in east Delhi.


الموضوعات
Bacterial Proteins/metabolism , Bacteriological Techniques/methods , Coenzymes/metabolism , Drug Resistance, Multiple, Bacterial , Enterobacteriaceae/enzymology , Enterobacteriaceae/isolation & purification , Enterobacteriaceae Infections/microbiology , Hospitals , Immunoassay/methods , India , Zinc/metabolism , beta-Lactamases/metabolism
6.
Indian J Med Microbiol ; 2011 Jul-Sept; 29(3): 249-253
مقالة ي الانجليزية | IMSEAR | ID: sea-143825

الملخص

Purpose: The newly emerging form of the so-called New Delhi Metallo-beta-lactamases (NDM-1) has been reported recently from patients worldwide and broadly thought as a potential source for the major global health problem. Thus, it is important to study the epidemiology of the so-called NDM-1 harbouring bacteria to prevent its further spread and to place effective control measures. The present study describes the use of the real-time polymerase chain reaction (PCR) assay for the detection of the bla NDM-1 gene using TaqMan probes among clinical isolates. Materials and Methods: Clinical isolates of Escherichia coli (11 strains), Klebsiella pneumoniae (17 strains) and Acinetobacter baumannii (six strains) that were resistant to either of the carbapenems (meropenem or imipenem) were included in the study. The presence of carbapenemases in such strains was confirmed using the modified Hodge test. A real-time PCR assay was optimized for the detection of NDM-1 using a cloned synthetic gene fragment followed by testing of the clinical isolates. The findings were further confirmed using PCR and gene sequencing. Results: TaqMan probe assay displayed a good detection limit with analytical sensitivity of the assay up to 10 copies of bla NDM-1 gene per reaction. The isolates of E. coli and K. pneumoniae revealed narrow range crossing point values (Cp values) between (12-17) cycles (mean Cp value 14), indicating number of bla NDM-1 gene copies of 106-108. The wider range of Cp values (15-34) cycles with a higher mean Cp value (23.6) was observed in A. baumannii with number of bla NDM-1 gene copies of 103-108. Conclusions: The study demonstrates that real-time PCR assay based on TaqMan chemistry is a useful technique for the detection of bla NDM-1 harbouring clinical isolates of E. coli, K. pneumoniae and A. baumannii. The assay has great precision in measuring the number of bla NDM-1 gene copies per specimen of DNA.


الموضوعات
Acinetobacter baumannii/drug effects , Acinetobacter baumannii/genetics , Acinetobacter baumannii/isolation & purification , Anti-Bacterial Agents/metabolism , Bacteriological Techniques/methods , Carbapenems/metabolism , Escherichia coli/drug effects , Escherichia coli/genetics , Escherichia coli/isolation & purification , Genes, Bacterial , Humans , Klebsiella pneumoniae/drug effects , Klebsiella pneumoniae/genetics , Klebsiella pneumoniae/isolation & purification , Real-Time Polymerase Chain Reaction/methods , beta-Lactam Resistance , beta-Lactamases/genetics , beta-Lactamases/metabolism
7.
Indian J Med Microbiol ; 2010 Oct-Dec; 28(4): 363-365
مقالة ي الانجليزية | IMSEAR | ID: sea-143740

الملخص

The aim of this study was to evaluate a nitrate reductase assay (NRA) for the direct detection of multidrug resistance (MDR) in Mycobacterium tuberculosis from 100 smear-positive sputum samples. The NRA results were compared with the reference proportion method for 100 sputum specimens for which comparable results were available. NRA results were obtained at day 7 for 61 specimens, results for 26 specimens were obtained at day 10, and the results for 13 specimens were obtained at day 14. Thus, 87% of NRA results were obtained in 10 days. NRA is a rapid, accurate, and cost-effective method for the detection of MDR in M. tuberculosis isolates as compared to the proportion method, which is time consuming. Therefore, NRA constitutes a useful tool for detection of tuberculosis drug resistance in low-resource countries with limited laboratory facilities due to its low-cost, ease of performance and lack of requirement of sophisticated equipment.

9.
مقالة ي الانجليزية | IMSEAR | ID: sea-135860

الملخص

Background & objectives: Protein energy malnutrition (PEM), an important cause of secondary immune deficiency, is associated with several abnormalities in the immune system including cytokine production. In the present study cytokine levels (both pro- and anti-inflammatory) were evaluated in protein energy malnourished children following nutritional rehabilitation with curd (Indian dahi) and leaf protein concentrate (LPC). Methods: Eighty moderately and severely malnourished children, 1-5 yr of age, received the WHO recommended diet for severe malnutrition, modified according to local dietary habits, containing in addition either curd or micronutrient-rich leaf protein concentrate, for a period of 15 days. Cytokine levels [tumour necrosis factor α (TNFα), interferon γ (IFNγ), interleukin-10 (IL-10), interleukin-4 (IL-4)] were measured before and after dietary rehabilitation. Results: The baseline cytokine levels (TNFα, IFNγ, IL-10 and IL-4) were high in malnourished children. Both the diets caused an increase in serum pro-inflammatory (TNFα, IFNγ), and anti-inflammatory (IL-10) cytokine levels after nutritional rehabilitation. The increase in IL-10 was significant in children receiving curd. There was an insignificant fall in IL-4 levels with both the diets. The cytokine response was comparable in children with moderate and severe malnutrition, as also in children < 2 yr to those between 2-5 yr. Interpretation & conclusions: The study suggests that cytokines (TNFα, IFNγ, IL-10 and IL-4) may serve as biological markers to assess the effect of functional foods like curd or LPC on immunity in malnutrition. Curd may help to maintain the balance in cytokine production by increasing the production of IL-10, and may be considered in place of milk in the nutritional rehabilitation of malnourished children.


الموضوعات
Child, Preschool , Cytokines/immunology , Dairy Products , Dietary Proteins/administration & dosage , Dietary Supplements , Humans , Infant , Nutritive Value , Plant Leaves/chemistry , Probiotics/administration & dosage , Protein-Energy Malnutrition/diet therapy , Protein-Energy Malnutrition/immunology , Random Allocation
10.
مقالة ي الانجليزية | IMSEAR | ID: sea-87189

الملخص

AIM: To determine the possibility of leptospirosis among patients from urban slums presenting with febrile illness during monsoon and post-monsoon season. METHODS: Evidence of leptospirosis in 180 patients with febrile illness was determined by looking for presence of immunoglobulin M (IgM) antibodies by leptospiral IgM enzyme linked immunosorbent assay (ELISA). The test was carried out on 160 Widal test negative and 20 Widal test positive sera received from febrile patients during June to September 2001. RESULTS: Twenty-seven out of 180 (15%) sera were positive for leptospiral IgM antibodies. CONCLUSIONS: This preliminary survey indicates that leptospirosis could be an important cause of febrile illness in patients from urban slums during monsoon and post-monsoon season.


الموضوعات
Fever/complications , Health Surveys , Humans , India/epidemiology , Leptospirosis/epidemiology , Poverty Areas , Seasons , Urban Population/statistics & numerical data
11.
Indian J Med Sci ; 2002 Jun; 56(6): 265-9
مقالة ي الانجليزية | IMSEAR | ID: sea-66740

الملخص

A comparative study involving SDS-PAGE of Salmonella typhi and other Bacteria was conducted. Protoplasmic antigens of Salmonella typhi. Salmonella paratyphi A, Salmonella typhimurium, Proteus sp, Klebsiellas sp. Pseudomonas aeruginosa, Shigella flexneri, Staphylococcus aureus were separated and compared on SDS-PAGE followed by checking of their cross reactivity by gel diffusion using antisera raised against whole cell and lysates of Salmonella typhi. Lines of identity between Salmonella typhi, Salmonella paratyphi A and Salmonella typhimurium were observed. No lines of identity were seen among Salmonella typhi, Pseudomonas aeruginosa and Sfaphylococcus aureus.


الموضوعات
Antigens, Bacterial/isolation & purification , Bacterial Proteins/immunology , Cross Reactions , Electrophoresis, Polyacrylamide Gel/methods , Immunodiffusion , Salmonella typhi/immunology
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