PubMed Health⌕ Search

Biomedical subjects

K K Mishra

Publications and source records attributed to K K Mishra.

10 recordsLinked to original sources

Antibiotic susceptibility of Helicobacter pylori clinical isolates: comparative evaluation of disk-diffusion and E-test methods.

Antimicrobial susceptibility of 25 Helicobacter pylori strains isolated from patients with acid peptic diseases were tested for in vitro sensitivity to commonly used antibiotics using disk-diffusion and E-test, methods. All strains tested were susceptible to tetracycline by E-test, with the minimum inhibitory concentration (MIC) values being <0.125 microg/ml for all strains except for 6 (<0.023 microg/ml). However 1 strain was resistant by disk-diffusion method. One strain was resistant to clarithromycin both by disk diffusion and E-test (MIC <48 microg/ml), and 1 strain was resistant only by disk diffusion. Only one strain was resistant to amoxicillin by disk diffusion and E-test (MIC >256 microg/ml). For ciprofloxacin, three strains were resistant by disk diffusion and two by E-test (MIC <32 microg/ml). Sixteen strains were resistant to metronidazole by disk diffusion and E-test (MIC >or= 8 microg/ml), and 1 was resistant only by E-test (MIC <48 microg/ml). Overall, 64% of the strains were resistant to metronidazole. The MIC for metronidazole was also tested by agar-dilution method, and metronidazole resistant strains had an, MIC >8 microg/ml. The disk-diffusion method showed excellent correlation with E-test results; there was 100% agreement for amoxicillin a other antibiotics showed 90% to 95% accuracy. Disk diffusion is cheaper than E-test (approximately 2.6 cents vs. 2.60 US dollars), is easy to perform, and is a reliable method for testing H. pylori susceptibility to antimicrobial agents in the clinical microbiology laboratory.

Anti-Bacterial Agents↗

UreC PCR based diagnosis of Helicobacter pylori infection and detection of cag A gene in gastric biopsies.

Polymerase chain reaction assay using ureC gene specific primers for the detection of Helicobacter pylori in gastric biopsy specimens from 116 dyspeptic patients was compared with other routine invasive diagnostic methods (culture, rapid urease test [RUT] and histology). In parallel, gastric biospy specimens from 54 patients and their corresponding Helicobacter pylori isolates were subjected to PCR with cagA targeting primers using standard protocols. Helicobacter pylori were detected in 53%, 43%, 48% and 50% of patients by PCR, RUT, culture and histological examination respectively. Based on histology and culture positive and at least three test positive result, 44 (37%), 46 (39%) and 26 (22%), and 56 (48%), 52 (44%) and 8 (6%) patients were classified as Helicobacter pylori positive, negative and indeterminate respectively. The sensitivity and specificity of PCR assay was the highest-95% and 100% when compared with both culture and histology positive, and at least any three positive results respectively. The result of cagA positivity in 54 gastric biopsy specimens and their corresponding Helicobacter pylori isolates were identical; 18 of 20 (90%) duodenal ulcer patients and 23 of 28 (82%) patients with chronic gastritis and 2 (40%) of 5 patients with portal hypertension and one gastric biopsy specimens from gastric cancer patients were found to be cagA positive. PCR-based method to detect Helicobacter pylori and the virulence gene cag A directly from gastric biopsy specimens appears to be promising and can curtail the lengthy process of culture-based approaches. The procedure proved to be rapid and reliable and could be utilized for diagnostic purposes.

Adult↗

Comparison of various conventional methods with a polymerase chain reaction assay for detecting methicillin-resistant & susceptible Staphylococcus aureus strains.

BACKGROUND & OBJECTIVES: Methicillin-resistant Staphylococcus aureus (MRSA), a major nosocomial pathogen world-wide, is often difficult to detect due to the heterogeneous nature of expression of oxacillin resistance. In the present study, various conventional methods were compared with polymerase chain reaction on 106 clinical isolates of Staph. aureus for detection of oxacillin resistance. METHODS: A total of 106 clinical isolates of Staph. aureus were tested for oxacillin resistance by disc diffusion, screen agar plates (3 micrograms and 6 micrograms/ml of oxacillin), oxacillin broth (3 micrograms/ml) and mecA based PCR. RESULTS: PCR detected mecA gene amplified product of 604 bp in 57 strains. Disc diffusion failed to detect 7 mecA positive strains but identified 5 mecA negative strains as oxacillin resistant. Screen agar 3 micrograms, screen agar 6 micrograms and oxacillin broth 3 micrograms detected 55, 53 and 55 respectively of the 57 mecA positive strains; however, they also falsely identified 5, 3 and 3 strains of mecA negative strains respectively as oxacillin resistant. The sensitivity, specificity and accuracy of disc diffusion, 3 micrograms screen agar, 6 micrograms screen agar and 3 micrograms oxacillin broth against PCR as gold standard were as follows: 87.7, 89.9 and 88.7 per cent; 96.5, 89.8 and 93.4 per cent; 93.0, 93.9 and 93.4 per cent; 96.5, 93.9 and 95.3 per cent respectively. INTERPRETATION & CONCLUSIONS: The present study demonstrated that disc diffusion test was least reliable and 3 micrograms broth had the highest sensitivity and specificity when compared with PCR for detection of oxacillin resistance. Because of variations among the methods, a combination of tests should be used for the accurate detection of MRSA till new guidelines by an appropriate body are formulated.

Base Sequence↗

A study on physiological changes in certain psychosomatic disorders with reference to cortisol, blood glucose and lipid profile.

Plasma cortisol, blood glucose, serum lipids and lipoproteins were estimated in diseased human subjects and normal control volunteers. Serum triglyceride (Tg) total cholesterol (Tc) and cholesterol content of very low density lipoprotein (VLDLc), low density lipoprotein (LDLc) and high density lipoprotein (HDLc) were assayed under lipid profile. Clinical investigations were carried out on 115 subjects which involved 30 control, 25 irritable bowel syndrome (IBS), 30 bronchial asthma and 30 rheumatoid arthritis patients. The results of this preliminary study showed a significant change in the levels of all the biochemical parameters in diseased subjects in comparison with controls. Increased levels of atherogenic lipids, Tg, VLDLc and LDLc were found in rheumatoid arthritis subjects. This suggests that arthritis subjects are relatively at higher risk of developing coronary heart disease. Furthermore hypercholesterolemia may aggravate the risk condition in arthritis patients by artereosclerosis. The significant elevation in the levels of plasma cortisol reveals the fact that rheumatoid arthritis is a stabilized and chronic psychosomatic disorder, since, homeostatic competence is disrupted following decline in the tendency of stress-response to return to normalcy.

Adult↗

A study on physiological changes in essential hypertension and rheumatoid arthritis with reference to the levels of cortisol, blood glucose, triglycerides and cholesterol.

The levels of plasma cortisol, blood glucose, serum triglycerides (TG) and total cholesterol (TC) were estimated in 175 human subjects (50 normal controls, 65 having essential hypertension and 60 suffering from rheumatoid arthritis. The results showed a significant elevation in the levels of plasma cortisol and blood glucose in both the stressed clinical groups with respect to controls. Increased levels of atherogenic lipids (TG and TC) were also observed in diseased group. However, in rheumatoid arthritis the biochemical changes were comparatively more pronounced than in hypertensives. The findings in vitro reveal that rheumatoid arthritis is a relatively more chronic and late onset disorder, since the functional performance of hypothalamo-pituitary-adrenocortical (HPA) axis declines with chronicity and the efficacy of adrenocortical response to return to normalcy becomes impaired.

Adult↗

Modulation and recovery of immune response of BALB/c mice to Shigella dysenteriae antigens after cyclophosphamide treatment.

We have analysed the suppressive effect of cyclophosphamide, when given in vivo, on the antibody response of BALB/c mice against Shigella dysenteriae antigens using ELISA and immunoblot. Out of various protocols tried, it was found that co-injection of cyclophosphamide at 150 mg/kg bodyweight, i.p., at the time of antigen administration and then after a lapse of 24 h during both primary and secondary immunizations, was the most effective in suppressing antibody response of mice. Analysis of sera by ELISA demonstrated the presence of some antibodies to S. dysenteriae antigens after secondary immunization, but immunoblot analysis using the same sera revealed complete suppression of antibody response. Animals whose antibody response was almost completely suppressed after two immunizations with co-injection of cyclophosphamide, when immunized again after the lapse of 14 days from the date of secondary immunization with Shigella antigens but without administration of cyclophosphamide, partially recovered their ability to respond to the same antigens. This protocol can now be used in mice to analyse the hierarchy of immunogenic epitopes present in a complex mixture of antigens.

Animals↗

Serological investigations on Indian kala-azar.

Detailed serological investigations were carried out in forty-nine active kala-azar (KA) cases in North Bihar, India. Various classes of immunoglobulin (IgG, IgA, and IgM) and third component of complement (C3) levels were determined in these sera and results were compared with those obtained in normal controls. Antibody titres were determined by the indirect haemagglutination (IHA) method using soluble Leishmania antigen. Immunoglobulin G and M class-specific antibody titres were also determined separately by the enzyme-linked immunosorbent assay (ELISA) method. Polyclonal hypergammaglobulinaemia with marked increase in serum IgG (and to a lesser extent in IgM) level was a rather common feature in the majority of these sera. Much of this immunoglobulin increase, however, appeared to be non-specific in nature and no absolute correlation could be noted between serum IgG or IgM levels and corresponding IgG or IgM antibody titres. Significant decrease in serum C3 level was observed in KA and this decrease was found to be independent of immunoglobulin levels or specific antibody titres. A fairly good correlation between aldehyde test and serum IgG level was evident from this study. Aldehyde-positive KA sera usually gave higher antibody titres than aldehyde-negative ones. Anti-leishmanial antibodies belonged mostly to IgG class although some IgM antibodies were also demonstrable. The latter class of antibodies probably appeared early in KA infection although their serological specificity was poorer to IgG antibodies. Out of forty-nine KA sera examined in this study thirty-six (73.5%) gave positive IHA titres while forty-six (93.9%) were positive by IgG-ELISA which appeared to be a highly specific and sensitive serodiagnostic method particularly for the early detection of KA cases.

Antibody Formation↗