Biomedical subjects

C N Paramasivan

Publications and source records attributed to C N Paramasivan.

At least 37 records · Page 2Linked to original sources

PubMed2003

Seroprevalence of human immunodeficiency virus (HIV) infection among tuberculosis patients in Tamil Nadu.

BACKGROUND & OBJECTIVES: The dual epidemic of HIV and tuberculosis is a cause for concern in those countries where these two infections are prevalent in epidemic proportions. We undertook a survey at two sites in North Arcot district of Tamil Nadu in 1992-1993, to know the seroprevalence of HIV infection among tuberculosis patients. The objective of this study was to re-examine the prevalence of HIV infection among tuberculosis patients in a repeat survey. METHODS: The study was undertaken in four centres: District Tuberculosis Centre (DTC), Vellore, Tuberculosis Sanatorium, Pennathur (Vellore), District TB Centre (DTC), Kancheepuram and the Government Thiruvotteswarar Tuberculosis Hospital (GTTH), Chennai in the northern part of Tamil Nadu during 1997-1998. A total of 2361 newly diagnosed TB patients were registered in this study. HIV serology after pre-test counseling was done along with sputum examination for acid-fast bacillus by smear and culture for mycobacteria for all patients. RESULTS: The overall HIV seroprevalence among TB patients was 4.7 per cent. The highest HIV seropositivity rate was found among patients aged 30-39 yr (10.6%). HIV seroprevalence showed a wide variation among the different centres ranging from 0.6. per cent in DTC, Kancheepuram to 9.4 per cent in Pennathur Sanatorium, Vellore. Sputum smear positivity was 88 per cent among the HIV-negative and 83 per cent among HIV-positive tuberculosis patients. INTERPRETATION & CONCLUSION: HIV infection is on the rise among TB patients in Tamil Nadu. Acid-fast smear microscopy is adequate for the diagnosis of pulmonary tuberculosis, and drug resistance among HIV positive patients is not a major problem at this point of time; hence antituberculosis regimens recommended by the Revised National Tuberculosis Control Program (RNTCP) can be used to treat HIV positive patients with tuberculosis.

Adult↗

PubMed2002

Surveillance of drug resistance in tuberculosis in two districts of South India.

SETTING: Tuberculosis Research Centre, Chennai, India. OBJECTIVES: To determine the levels of drug resistance in new and previously treated cases of pulmonary tuberculosis in the composite districts of North Arcot (Tamil Nadu State) and Raichur (Karnataka State) in South India. DESIGN: Two specimens of sputum from 320 patients attending 23 participating centres in North Arcot district and 314 patients from 20 participating centres in Raichur district were tested for drug susceptibility to isoniazid (H), rifampicin (R), ethambutol (E), streptomycin (S) and ofloxacin (O). The studies were undertaken using the guidelines prescribed by the WHO/IUATLD Working Group on Anti-tuberculosis Drug Resistance Surveillance. RESULTS: In North Arcot district, resistance to any drug tested was found in 27.7% of new cases; any H resistance in 23.4%, any R resistance in 2.8% and multidrug resistance (resistance to at least H and R: MDR) in 2.8%. In previously treated cases, resistance to any drug was observed in 81.2%, and any resistance to H, R and HR in 81%, 69% and 69%, respectively. In Raichur district, resistance to any drug was observed in 21.9% of new cases; any resistance to H, R and HR (MDR) was found in 18.7%, 2.5% and 2.5%, respectively. All previously treated patients were resistant to H and R (100%). CONCLUSION: In North Arcot district, the proportion of MDR-TB in newly diagnosed patients has marginally increased over the last 10-15 years, whereas it has remained fairly constant in Raichur district. An increase in resistance was noted in previously treated cases in both districts, although the numbers of patients are small.

Antitubercular Agents↗

PubMed2001

Use of vancomycin in the culture of Mycobacterium tuberculosis from gastric lavage.

BACKGROUND & OBJECTIVES: Earlier studies from the Tuberculosis Research Centre, Chennai, on culture of Mycobacterium tuberculosis from gastric lavage (GL) specimens in selective Kirchner's medium (SK) resulted in a loss of 60 per cent culture results due to contamination with aerobic spore bearers (ASB). Addition of vancomycin to SK (SKV) effectively reduced the contamination rate to 20 per cent. The objective of the present study was to further reduce the contamination by collecting the specimens in bottles containing vancomycin, thus providing continuous exposure of the sample to the drug, which is bactericidal to ASB. METHODS: One thousand GL specimens collected from children in vancomycin containing bottles were decontaminated and cultured in SK medium, with and without vancomycin, subcultured on Lowenstein Jensen (LJ) medium and the culture results compared. RESULTS: The contamination of cultures in SK and SKV was 15 and 4 per cent respectively when the specimens were collected in bottles containing vancomycin compared to 60 and 20 per cent contamination reported in the earlier studies. INTERPRETATION & CONCLUSION: The reduced contamination in SK and SKV is most likely due to the collection of samples in vancomycin containing bottles. Although a concurrent comparison of samples processed in vancomycin free conditions would have been ideal, it could not be done due to practical difficulties. The study thus confirms the value of vancomycin as a major deterrent for contamination due to aerobic spores and better results can be obtained if vancomycin is used in sample collection bottles, transport media and liquid culture media used in mycobacteriology laboratories particularly in humid and tropical environment.

Culture Media↗

PubMed2001

Comparison of different methods of assessing in vitro resistance of Mycobacterium tuberculosis to rifampicin.

BACKGROUND & OBJECTIVES: Definitions of in vitro resistance to rifampicin in strains of Mycobacterium tuberculosis by different methods have not been consistent, leading to variations in the interpretation and validity of results. This study compared three methods of defining in vitro resistance to rifampicin. METHODS: (i) A total of 598 clinical isolates of M. tuberculosis were concurrently compared by the minimal inhibitory concentration (MIC) and the proportion method on Lowenstein-Jensen medium; (ii) 54 strains tested by the MIC method were retested by the proportion method and the BACTEC radiometric method; and (iii) 72 strains which yielded an MIC of 64 mg/l by the MIC method were retested by the same method. RESULTS: Out of 598 cultures tested by the MIC and the proportion methods, identical classification as susceptible or resistant was observed in 99.7 per cent. A 100 per cent agreement was observed when 54 strains were tested by the MIC, proportion and BACTEC radiometric methods. When 72 strains with an MIC of 64 mg/l were retested by the same method, 61 (85%) yielded a lower MIC, 9 (12%) gave the same MIC while 2 (3%) yielded a higher MIC of 128 mg/l, reflecting perhaps the inherent limitations of the variations in the inoculum size. INTERPRETATION & CONCLUSION: All 3 definitions of resistance, viz., an MIC of 128 mg/l, a proportion of 1 per cent or more on 40 mg/l by the proportion method, both on L-J medium and a growth of 1 per cent or more on 2 mg/l by the radiometric method were found to be equally satisfactory.

Antibiotics, Antitubercular↗

PubMed2000

A multicentre study of the early bactericidal activity of anti-tuberculosis drugs.

The early bactericidal activities (EBAs) of 300 mg isoniazid, 18.5 mg isoniazid, 600 mg rifampicin and 800 mg ofloxacin given daily to 262 patients with newly diagnosed pulmonary tuberculosis in Cape Town, Nairobi, Madras and Hong Kong were measured by counting cfu and total acid-fast bacilli in sputum collections taken pre-treatment (S1), at 2 days (S3) and at 5 days (S6). In Cape Town, Nairobi and Madras, the cfu findings suggested that isoniazid produced a massive kill, perhaps of actively growing organisms, during the first 2 days (mean S1-S3 EBAs of 0.636-1.006) but was almost inactive thereafter (mean S3-S6 EBAs of 0.000-0.081), whereas rifampicin maintained moderate activity against slowly growing organisms throughout the 5 days (mean S3-S6 EBAs of 0.242-0.305). This finding suggests that EBAs measured during the 2-5 day interval might be able to assess the sterilizing activity of drugs. Ofloxacin had moderately high mean S1-S3 EBAs of 0.130-0.391. However, in Hong Kong rifampicin appeared to be the most bactericidal drug from the start, possibly because patients had more chronic disease. A method of adjusting the cfu EBAs using total counts was devised which decreased the variability between patients within a treatment group without altering the mean cfu EBA. This resulted in a large gain in precision in Hong Kong, suggesting that their estimates were greatly affected by type II variation, due to dilution of pus by saliva and bronchial secretions, whereas small or no gains were obtained in the other three centres, suggesting that the main cause of variability was type I, due to other factors.

Adult↗

PubMed2000

Risk of development of tuberculosis in HIV-infected patients.

OBJECTIVE: To estimate the rate of development of active tuberculosis (TB) in a cohort of human immunodeficiency virus (HIV) positive patients, and to identify the characteristics of these patients. DESIGN: A total of 175 HIV-positive individuals were recruited from clinics for sexually transmitted diseases and followed up for 31 +/- 6.8 months. Clinical examination, chest X-ray, sputum smear for acid-fast bacilli and culture for mycobacteria and HIV serology were performed at the time of registration and repeated periodically. RESULTS: Seventeen patients had TB at intake and another 24 developed TB during follow-up, giving a breakdown rate of 6.9/100 person-years (p-y) (95% confidence interval [CI] 4.1-9.6). The attack rates were similar in tuberculin positive (7.1/100 p-y, 95%CI 3.4-10.8) and negative (6.7/100 p-y, 95%CI 2.6-10.8) patients. There was a trend towards higher mortality in patients who developed TB (10.5/100 p-y, 95%CI 4.8-15.2) compared to those who did not (6.1/100 p-y, 95%CI 3.2-8.8). CONCLUSIONS: The results of this study provide information regarding the high risk of development of active tuberculosis and its associated mortality in HIV-infected persons. The risk of developing TB appears to be equally high in tuberculin positive and negative individuals, suggesting that new infections could play a major role in this susceptible population.

AIDS-Related Opportunistic Infections↗

PubMed1999

Associations of HLA-DRB1, DQB1 and DPB1 alleles with pulmonary tuberculosis in south India.

SETTING: Tuberculosis is endemic in south India: sputum positive pulmonary tuberculosis is predisposed by HLA-DR2 in south India and few other populations of the world. OBJECTIVE: To study HLA-DRB1, DQB1, DQA1 and DPB1 allelic polymorphism in pulmonary tuberculosis patients and endemic controls from south India. DESIGN: One hundred and twenty-six, sputum positive pulmonary tuberculosis patients and 87, endemic controls, from Madurai were studied for MHC class II allelic polymorphism by PCR-SSOP method. XI IHWC primers and probes and non-radioactive probing methods were employed. RESULTS: HLA DRB1*1501 and DQB1*0601 predisposed for pulmonary tuberculosis (DRB1*1501: odds ratio (OR) = 2.68, 95% confidence interval (CI) = 1.30-5.89, P value (P) = 0.013, aetiological fraction (EF) = 0.17; DQB1*0601: OR = 2.32, CI = 1.29-4.27, P = 0.008, EF = 0.26). Haplotype DRB1*1501-DQB1*0601 was higher in patients (1324 per 10,000, X2 = 27.07) than controls (F = 404/10,000, X2 = 8.84). In a subset of 63 caste matched samples, DPB1*04 was preventive (OR = 0.45, CI = 0.21-0.95, P = 0.036, PF = 0.26): the distributions of DRB1*1501-DQB1*0601-DPB1*04 phenotypes were different between patients and controls (P = 0.0092). These alleles were predominant in patients and controls of T5SU caste. CONCLUSION: HLA-DRB1*1501 and DQB1*0601 predisposed to sputum positive pulmonary tuberculosis, and DPB1*04 was preventive and epistatic to this risk. Caste T5SU is an ideal model to study immunology of tuberculosis.

Adult↗

PubMed1999

A histological spectrum of host responses in tuberculous lymphadenitis.

A total of 446 lymph node biopsy specimens showing histological evidence of tuberculosis were classified into four groups based on the organization of the granuloma, the type and numbers of participating cells and the nature of necrosis. These were, hyperplastic (22.4%)--a well-formed epithelioid cell granuloma with very little necrosis, reactive (54.3%)--a well-formed granuloma consisting of epithelioid cells, macrophages, lymphocytes and plasma cells with fine, eosinophilic caseation necrosis, hyporeactive (17.7%)--a poorly organized granuloma with macrophages, immature epithelioid cells, lymphocytes and plasma cells and coarse, predominantly basophilic caseation necrosis and nonreactive (3.6%)--unorganized granuloma with macrophages, lymphocytes, plasma cells and polymorphs with non caseating necrosis. Though the number of bacilli in the sections differed in each group, there were no differences in culture positivity, Mantoux reaction or the clinical features. It is likely that the spectrum of histological responses seen in tuberculous lymphadenitis is the end result of different pathogenic mechanisms underlying the disease.

Adolescent↗

PubMed1999

Early results from indirect drug susceptibility test for tubercle bacilli.

The indirect susceptibility test results on L-J medium for tubercle bacilli against streptomycin, isoniazid and rifampicin were read at the end of 2 wk and compared with the results at 4 wk. It was found that drug resistance could be correctly predicted in over 70 per cent of cultures including multi-drug resistant tuberculosis (MDR TB) strains at the end of 2 wk. The susceptibility to para-nitrobenzoic acid (PNB) read at 2 wk was able to distinguish non-tuberculous mycobacteria from Mycobacterium tuberculosis cultures. The early detection of resistance by this procedure requires only minimum inputs, and can benefit the majority of patients harbouring drug resistant tubercle bacilli.

Antitubercular Agents↗

PubMed1999

Evaluation of various methods of susceptibility to ofloxacin in strains of Mycobacterium tuberculosis.

A comparison of three methods of susceptibility testing was undertaken on 30 susceptible and 25 resistant strains of Mycobacterium tuberculosis to determine an acceptable in vitro definition of resistance of ofloxacin. The strains were tested by the proportion method on Lowenstein Jensen (L-J) and 7H11 media and also by the BACTEC radiometric method. Using a criterion of 1 per cent or more growth at a concentration of 2 mg/1, there was a 100 per cent agreement with the conventional MIC method by the proportion tests on L-J as well as on 7H11 media. The BACTEC radiometric method, at the same concentration, yielded 98 per cent agreement. Thus, any of these methods could be used depending upon the infrastructure available.

Anti-Infective Agents↗

PubMed1998

Bioluminescence assay of adenosine triphosphate in drug susceptibility testing of Mycobacterium tuberculosis.

Twenty three clinical isolates M. tuberculosis and the reference strain, M. tuberculosis H37Rv were tested for their susceptibility to trifluoperazine (TFP) by the standard broth dilution method and the bioluminescence assay. The results showed that in 15 of the 23 isolates, the minimal inhibitory concentration (MIC) was identical in both the methods and in the remaining 8 isolates the difference in the MIC values between the methods, was less than two fold and was not significant. The findings suggest that the measurement of adenosine triphosphate (ATP) by bioluminescence assay can be employed as an alternative method for the rapid screening of clinical isolates for their susceptibility to anti-mycobacterial agents.

Adenosine Triphosphate↗

PubMed1998

Action of metronidazole in combination with isoniazid & rifampicin on persisting organisms in experimental murine tuberculosis.

To study the activity of metronidazole on persisting tubercle bacilli, BALB/c mice were infected with Mycobacterium tuberculosis and, after 14 days, treated with isoniazid (H) or rifampicin (R) or isoniazid + rifampicin (HR) for 2 months. An untreated group and a group treated with metronidazole (M) alone served as controls. At the end of 2 months, M was added to the H, R, and HR regimen in half the mice, and the treatment was continued for 1 more month in all mice. At the end of treatment, no viable organisms were detected in the lung or spleen of mice treated with HR or HRM regimens. In contrast, compared to the mice treated with R alone, the log10 colony forming units (cfu) of mice treated with RM were lower by 1.84 and 0.52 in the lung and spleen, respectively. Similarly, compared to the H group, the log10 cfu were lower by 0.67 in the spleen of mice treated with HM, and no additional effect due to M was seen in the lung. Three months after stopping treatment, viable organisms were isolated from both the organs of all the groups. However, the log10 cfu in the lung and spleen for the groups with metronidazole were below the log10 cfu for the respective single or 2 drug groups, except the log10 cfu in the lung for the RM group. These findings suggest that metronidazole, given with bactericidal drugs such as rifampicin and isoniazid may be of value in eliminating persisting tubercle bacilli, but further studies are warranted.

Animals↗

PubMed1998

Evaluation of the BACTEC radiometric method in the early diagnosis of tuberculosis.

A comparison of the BACTEC radiometric method with the conventional culture and drug susceptibility testing methods on isolates from clinical specimens in pulmonary and extrapulmonary tuberculosis, childhood TB and TB in HIV-infected individuals was undertaken. In the case of pulmonary TB, the rate of isolation of positive cultures was significantly faster with the BACTEC method, with 87 per cent of the positives being obtained by 7 days, and 96 per cent by 14 days. However, while there was no difference in the total number of positive cultures by the two methods in smear positive pulmonary tuberculosis, in smear negative pulmonary TB, the BACTEC method yielded more number of positive cultures. In extrapulmonary TB, HIV-TB and childhood TB, although the BACTEC method did not yield additional positives, the detection of positives was considerably faster than by the conventional methods, in which the degree of growth was also scanty. The agreement in drug susceptibility tests was 94 per cent for streptomycin and isoniazid, 99 per cent for rifampicin and 91 per cent for ethambutol. Further, most of the drug susceptibility test results became available within 8 days by the BACTEC method. By facilitating early diagnosis, the BACTEC method may prove to be cost effective in a population with a high prevalence of tuberculosis, particularly in the extrapulmonary and paucibacillary forms of the disease.

Carbon Radioisotopes↗

PubMed1997

Transportation of lymph node biopsy specimens in selective Kirchner's liquid medium for culture of tubercle bacilli.

Lymph node biopsy specimens, obtained from 297 paediatric and adult patients with tuberculous lymphadenitis at Madurai, were transported in selective Kirchner's liquid medium (KL-T) to the Tuberculosis Research Centre, Madras and processed for culture. Mycobacterium tuberculosis was isolated from 201 (68%) specimens. Of the 192 specimens received within 4 days of resection, 134 (69.8%) yielded M. tuberculosis on culture and of the 105 specimens received after 5 days, 67 (63.8%) were culture positive; the difference was not statistically significant. By incubating KL-T alone further, after removing the gland for processing, it was found that mere contact with the excised node during transportation was enough to retrieve 77 (38.3%) of the total of 201 positive isolates obtained, the delay did not affect the culture positivity rate. Thus, lymph node specimens for culture of tubercle bacilli can be stored in the refrigerator for up to 15 days and transported in KL-T at ambient temperature for 18-20 h without any loss in culture positivity.

Adult↗

PubMed1997

Molecular characterization of Mycobacterium avium complex isolates giving discordant results in AccuProbe tests by PCR-restriction enzyme analysis, 16S rRNA gene sequencing, and DT1-DT6 PCR.

Based on cultural and biochemical tests, a total of 84 strains (72 clinical and 12 environmental isolates from the Caribbean Isles, Europe, and the Indian subcontinent) were identified as members of the Mycobacterium avium complex (MAC). They were further characterized with MAC, M. avium, and M. intracellulare probes of the AccuProbe system, and this was followed by selective amplification of DT6 and DT1 sequences. Seventy isolates gave concordant results; 63 were identified as M. avium, 5 were identified as M. intracellulare, and 24 remained untypeable by both methods. Fourteen isolates gave discrepant results, as they were DT1 positive but gave negative results by the M. intracellulare AccuProbe test. Consequently, a detailed molecular analysis of all DT1-positive isolates (14 discrepant strains plus 5 M. intracellulare strains) was performed by PCR-restriction analysis (PRA) of the hsp65 gene and 16S rRNA gene sequencing. The results confirmed the reported heterogeneity of M. intracellulare, as only 6 of 19 isolates (32%) gave PRA results compatible with published M. intracellulare profiles while the rest of the isolates were grouped in four previously unpublished profiles. 16S rRNA gene sequencing showed that only 8 of 19 isolates (42%) were related to M. intracellulare IWGMT 90247 (EMBL accession no. X88917), the rest being related to MCRO19 (EMBL accession no. X93030) and MIWGTMR10 (EMBL accession no. X88915). In conclusion, we have characterized a significant number of MAC isolates which were not identified by the AccuProbe test, PRA, or 16S rRNA sequencing. However, all of them were identifiable by DT1-DT6 PCR (they were DT6 negative and DT1 positive) and could be tentatively identified as M. intracellulare based on previously published observations. It is noteworthy that the majority of such isolates (14 of 19) were from the Indian subcontinent, with 12 of 14 being environmental isolates. Our study confirms the marked heterogeneity of M. intracellulare isolates and shows the utility of in-house DT1 PCR to detect this group of isolates, which would otherwise have been missed by the AccuProbe system in a routine clinical microbiology laboratory.

Base Sequence↗

PubMed1997

In vitro susceptibility of clinical isolates of Mycobacterium tuberculosis to cefadroxil--a cephalosporin antibiotic.

The bactericidal activity (BA) of cefadroxil, a semisynthetic cephalosporin antibiotic, against M. tuberculosis H37Rv was studied in Middlebrook 7H9 medium. Cefadroxil showed good BA (average fall of viable counts = log10 0.32 colony forming units/ml/day) against the log phase culture of M. tuberculosis H37Rv. Its minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were found to be 15 micrograms/ml or less. The MIC of cefadroxil for 29 clinical isolates of M. tuberculosis and a laboratory strain, M. tuberculosis H37Rv was also determined by agar dilution method using Middlebrook 7H11 agar as a screening procedure. The MIC of cefadroxil was found to be 10 micrograms/ml or less for M. tuberculosis H37Rv and 16 (55.1%) of 29 clinical isolates tested. The MIC for 3 of 10 drug sensitive and 9 of 19 drug resistant isolates was 40 or more, a concentration much higher than the peak plasma concentration (28 micrograms/ml) attained in human beings. The higher MIC observed in 12 of 29 clinical isolates irrespective of their susceptibility pattern requires further studies to assess the usefulness of cefadroxil in the treatment of tuberculosis.

Cefadroxil↗

PubMed1997

Minimal inhibitory concentrations of sulbactam/ampicillin against drug sensitive and drug resistant isolates of Mycobacterium tuberculosis.

A total of 92 isolates of Mycobacterium tuberculosis consisting of equal numbers of sensitive and resistant strains was tested for their susceptibility to sulbactam and ampicillin (in the ratio of 1:2) on Lowenstein-Jensen (LJ) and 7H11 agar media. The geometric mean MIC was 63.97 micrograms/ml for the drug sensitive strains and 65.92 micrograms/ml for the resistant strains, and the overall mean was 65.01 micrograms/ml. The high MIC on LJ medium could be attributed to the higher protein content which resulted in greater binding of sulbactam/ampicillin. On the other hand, the geometric mean MIC on 7H11 medium was 26.73 micrograms/ml for sensitive strains and 23.82 micrograms/ml for resistant strains; the overall mean being 25.23 micrograms/ml. Although these MICs of sulbactam-ampicillin are higher than those reported earlier, they can be easily achieved in serum. Further studies on experimental tuberculosis and in humans will be needed to prove the efficacy of sulbactam/ampicillin in the treatment of patients with multidrug resistant tuberculosis.

Ampicillin↗

PubMed1997

Minimal inhibitory concentrations of lomefloxacin and minocycline against drug-sensitive and drug-resistant isolates of M. tuberculosis compared on L-J and 7H11 media.

The in vitro activity of lomefloxacin and minocycline was tested against 46 strains of M. tuberculosis resistant to streptomycin (S), isoniazid (H) and rifampin (R) or SHR and 51 strains sensitive to SHR by the minimal inhibitory concentration (MIC) method on two different media, namely, Lowenstein-Jensen (L-J) and Middlebrook 7H11. The results of the study showed that, irrespective of the medium used, minocycline had little activity against the strains tested and the MIC was > 64 micrograms/ml. The MIC of lomefloxacin in 7H11 medium ranged from 2 to 16 micrograms/ml. There were highly significant differences in the MICs of lomefloxacin in L-J compared with 7H11. The results suggest that the activity of lomefloxacin against M. tuberculosis merits further study.

Anti-Infective Agents↗