Serum and cerebrospinal fluid concentrations of rifampicin at two dose levels in children with tuberculous meningitis.

Twenty patients of tuberculous meningitis (TBM) in age group of 6 months to 10 years included in the study were divided into two groups of 10 patients each. Rifampicin was administered in dosage of 10 mg and 7.5 mg/kg bw to each patient of groups I and II respectively. Drug concentrations in serum and CSF of these patients were measured by a microbiological tube dilution method using a strain of Sarcina lutea. In group I mean serum and CSF concentration was 3.84 micrograms/ml and 0.178 microgram/ml respectively, while in group II it was 2.16 micrograms/ml and 0.206 microgram/ml respectively. These concentrations were many times higher than the MIC against Mycobacterium tuberculosis. Mean percentage penetration of rifampicin in CSF was 5 and 10% in group I and II respectively. We recommend similar studies in large number of children before advocating the therapy with low dose of rifampicin in TBM.

Detection of IgM antibodies in pulmonary tuberculosis by ELISA using A60 antigen.

An enzyme immunosorbent assay (ELISA) was evaluated as a serodiagnostic test in patients with active tuberculosis (Group I), clinically suspected pulmonary tuberculosis (Group II) and pulmonary diseases other than tuberculosis and normal healthy subjects (Group III). and compared with culture and sputum smear examination results. ELISA was found to have a sensitivity of 77.5% and a specificity of 87.5%. Sputum smear had a sensitivity of 65% and specificity of 100%. In patients clinically suspected to have pulmonary tuberculosis ELISA had a sensitivity of 60%. Thus ELISA using A60 antigen can facilitate the diagnosis of tuberculosis in patients with active as well as suspected pulmonary tuberculosis and can provide results within hours of receipt of samples.

Initial and acquired drug resistance of Mycobacterium tuberculosis in east Delhi.

One hundred and fifty six strains of Mycobacterium tuberculosis, isolated from cases of pulmonary tuberculosis were subjected to sensitivity test to detect initial and acquired drug resistance to Streptomycin, isoniazid, Rifampicin, Ethambutol and Pyrazinamide. Initial and acquired drug resistance was observed to one or more drugs in 16% and 24.4% of the patients respectively. Strains resistant to Rifampicin and Ethambutol were resistant to Isoniazid also. Drug resistance was unrelated to age and sex of the patients.

Enterotoxin production and plasmid profiles in Salmonella typhimurium isolated from man.

One hundred twenty-six isolates of Salmonella typhimurium from various clinical sources were tested for enterotoxin production and characterization of plasmid profile. Cell-free culture supernates and polymyxin B extracts of all the strains were assayed by rabbit ileal loop and skin permeability tests. Enterotoxic activity was detected in culture supernates of 32 strains. Twenty-one strains by both rabbit ileal loop and skin permeability tests, nine strains by skin permeability test, and two strains by rabbit ileal loop test were positive. Live culture of three enterotoxic strains, positive in culture supernates produced ileal secretion. Polymyxin B extracts from 6 hours and 18 hours broth cultures of all the strains were devoid of enterotoxicity. Ileal mucosa exposed to culture supernate of enterotoxigenic strains showed swollen and blunted villi with submucosal oedema while those exposed to polymyxin B extracts showed shortening of villi and sloughing of epithelial lining. Plasmid profiles of enterotoxigenic strains were heterogenous and grouped into 20 different profiles. No correlation could be established between plasmid profile, R-pattern, and enterotoxin production.

Plasmid profile and phage type of Salmonella typhimurium strains encountered in different regions of India.

A total of 190 Salmonella typhimurium strains encountered in different parts of India were characterized on the basis of plasmid profile, phage type and antimicrobial resistance pattern. Recent trends in the epidemiology of R-plasmids were also studied. The majority of S. typhimurium strains (90.5%) were untypable by phage typing. Only 18 strains (9.5%) were phage typable. The phage untypable strains isolated from northern (57) central (65), and southern (50) regions of India could be subgrouped into 24, 12 and 16 different plasmid profiles respectively. Heterogeneity was the prominent feature although most of the plasmid profiles were related among strains isolated from particular place. A great diversity among small plasmids (2.7-8.3 kb) made subgrouping of majority strains (71%) with R-pattern ApCmKmSmSuTcTp possible. Conjugation studies and plasmid profile analysis of transconjugants revealed all the strains to harbour non conjugative non-auto transmissible plasmids with the exception of 7.2 and 2.7 kb plasmids which were not mobilizable.

Plasmid profiles, adhesive & invasive properties of Salmonella ser. Typhimurium strains: phage types 99 & 36.

Adhesive and invasive properties were compared with plasmid profile in S. Typhimurium strains of phage type 99(10) and 36(10). All strains of phage type 99 were multiple drug resistant (ApCmKmSmSuTcTp) and harboured small plasmids of 2.4-5.2 MDal. Six strains of the phage type 36 had ApCmTc R-pattern and two with only ampicillin resistance, carried plasmids of molecular size 2.6-5.2 MDal; two strains were sensitive to all antibiotics and devoid of plasmids. None of the strains were found to harbour high molecular weight plasmids. All plasmid positive strains of phage types 99 and 36 could be divided into two groups of three plasmid patterns each, which were phage type specific. All plasmid positive and negative strains adhered and invaded HeLa cells to different degrees. No correlation could be established between plasmid profile and adhesion invasion characteristics. High molecular weight plasmids therefore are unlikely to be essential for adhesion and invasion.

Plasmid profile analysis of Salmonella ser. Typhimurium phage untypable strains encountered in diverse geographic regions of India.

Plasmid profile analysis and antibiotic resistance pattern determination were carried out for 117 phage untypable S. Typhimurium strains. Majority of the strains (82%) were resistant to all the seven antibiotics tested, R-pattern being ApCmKmSmSuTcTp, rest (12%) showed heterogenous R-patterns. Plasmid DNA analysis revealed phage untypable strains to harbour large (58.8-114.3 MDal), intermediate size (36 MDal, 42 MDal) and small (1.8-5.2 MDal) plasmids with varying molecular weights. All the phage untypable strains could be subgrouped by plasmid profile analysis into 23 plasmid patterns. Plasmid profile analysis could discriminate large number of phage untypable strains on the basis of their plasmid pattern.

Disinfectant induced changes in the antibiotic sensitivity and phage typing pattern in Staphylococcus aureus.

Serial passage through disinfectants at sub-minimal inhibitory concentrations induced antibiotic resistances in 30.5% of the derived phenotypic variants of five strains of Staphylococcus aureus. A proportion of these phenotypic variants showed changes in their phage typing pattern by becoming susceptible to an increased number of phages, and a change from phage non-typable to typable was also observed. A loss of temperate phages was noted in some variants when subjected to reverse phage typing.

Isoenzyme studies of Giardia lamblia isolated from symptomatic cases.

Strains of Giardia lamblia were isolated from symptomatic cases of giardiasis and axenized in the laboratory. Electrophoretic mobility patterns of four enzymes, viz., EC 5.3.1.9 glucose phosphate isomerase (GPI); EC 1.1.1,4.0.L-malate; NADP+ Oxidoreductase (Oxaloacetate decarboxylating) (ME); EC 2.7.5.1 phosphoglucomutase (PGM); and EC 2.7.1.1 hexokinase (HK) of the lysates prepared from these isolates were studied using starch-gel. Based on differences in mobility patterns of PGM and HK, the four strains studied could be grouped into three different isoenzyme types (Zymodemes). ME mobility was identical in all the four strains. Some relative difference was seen in the mobility of GPI, though the pattern of mobility was similar in all the strains.

Immunodiagnosis of amoebiasis.

Serological studies were done on 127 cases using three different techniques namely indirect haemagglutination (IHA), indirect fluorescent antibody test (IFT) and counter immunoelectrophoresis (CIEP) to detect antiamoebic antibodies. All amoebic liver abscess cases showed significant titre of antibodies by all the three tests used. In the group of patients suffering from amoebic pathology of liver, 90.47 per cent were positive by IHA, 100 per cent by IFT and 85.71 per cent by CIEP respectively. Among amoebic dysentry and amoebic colitis cases 81.81 per cent and 80.64 per cent respectively were positive by IHA. The corresponding figures for IFT were 100 per cent and 74.19 per cent and for CIEP 90.90 per cent and 64.51 per cent respectively. Follow up study showed no significant fall in antibody titre in nine cases studied upto 10 weeks after treatment. Amoebic antigen could be detected in pus from all the nine cases with amoebic liver abscess by CIEP test.

Cellulose acetate precipitation test in sero-diagnosis of amoebiasis.

Cellulose acetate precipitation (CAP) test for the detection of antibody against Entamoeba histolytica using the axenic antigen, was performed on the 127 serum samples obtained from patients with amoebic liver abscess (14), amoebic hepatitis (21), amoebic dysentry (11), amoebic colitis (31), other parasitic infestations (25) and normal individuals (25). The percent positivity was 100, 95.23, 90.9, 67.74 and 16 and 12 respectively whereas the corresponding figures for the indirect immunoflourescence (IFAT) test were 100, 100, 100, 74.19, 12 and 8 respectively. Although CAP is not as good a test as IFAT, yet it can be recommended for routine testing due to its sensitivity, speed of performance and applicability to a single serum sample.