-
A Martin,
N Morcillo,
D Lemus,
E Montoro,
M A da Silva Telles,
N Simboli,
M Pontino,
T Porras,
C León,
M Velasco,
L Chacon, L Barrera,
V Ritacco,
F Portaels,
J C Palomino
[show abstract]
[hide abstract]
ABSTRACT: A multicentre evaluation was performed to assess two rapid low-cost methods, MTT (3-[4.5-dimethylthiazol-2-yl]-2.5-diphenyltetrazolium bromide) and resazurin assays, for testing the susceptibility of Mycobacterium tuberculosis to the first-line anti-tuberculosis drugs rifampicin (RMP), isoniazid (INH), ethambutol (EMB) and streptomycin (SM).
Thirty coded M. tuberculosis strains were sent to seven laboratories located in Latin America, representing six countries. Each site performed the colorimetric assays, MTT and resazurin, blind for the first-line drugs RMP, INH, EMB and SM. The minimum inhibitory concentration results obtained were compared to the conventional proportion method on Lowenstein-Jensen medium.
After establishing the breakpoint concentrations, excellent results were obtained for RMP, INH and EMB, with levels of specificity and sensitivity of between 96% and 99%.
MTT and resazurin assays are promising, accessible new alternative methods for middle- and low-resource countries that need low-cost methods to perform rapid susceptibility testing of M. tuberculosis to key anti-tuberculosis drugs.
The international journal of tuberculosis and lung disease: the official journal of the International Union against Tuberculosis and Lung Disease 09/2005; 9(8):901-6. · 2.73 Impact Factor
-
B López,
D Aguilar,
H Orozco,
M Burger,
C Espitia,
V Ritacco, L Barrera,
K Kremer,
R Hernandez-Pando,
K Huygen,
D van Soolingen
[show abstract]
[hide abstract]
ABSTRACT: In the last decade, an unprecedented genetic diversity has been disclosed among Mycobacterium tuberculosis strains found worldwide. However, well-conserved genotypes seem to prevail in areas with high incidence of tuberculosis. As this may be related to selective advantages, such as advanced mechanisms to circumvent [M. bovis Bacille Calmette-Guerin (BCG)-induced] host defence mechanisms, we investigated the influence of strain diversity on the course of experimental disease. Twelve M. tuberculosis strains, representing four major genotype families found worldwide today, and the laboratory strain H37Rv were each used to infect BALB/c mice by direct intratracheal injection. Compared with H37Rv, infections with Beijng strains were characterized by extensive pneumonia, early but ephemeral tumour necrosis factor-alpha (TNF-alpha) and inducible isoform of nitric oxide synthetase (iNOS) expression, and significantly higher earlier mortality. Conversely, Canetti strains induced limited pneumonia, sustained TNF-alpha and iNOS expression in lungs, and almost 100% survival. Strains of the Somali and the Haarlem genotype families displayed less homogeneous, intermediate rates of survival. Previous BCG vaccination protected less effectively against infection with Beijing strains than against the H37Rv strain. In conclusion, genetically different M. tuberculosis strains evoked markedly different immunopathological events. Bacteria with the Beijing genotype, highly prevalent in Asia and the former USSR, elicited a non-protective immune response in mice and were the most virulent. Future immunological research, particularly on candidate vaccines, should include a broad spectrum of M. tuberculosis genotypes rather than a few laboratory strains.
Clinical & Experimental Immunology 08/2003; 133(1):30-7. · 3.36 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Resistance of Mycobacterium tuberculosis to both isoniazid (INH) and rifampicin (RPM), the most important antituberculosis drugs, with or without simultaneous resistance to other drugs, is known as multidrug resistance (MDR). It is the main obstacle to attain the cure of patients by the specific treatment, and a threat to the tuberculosis control. Between 1994 and 1997, several Latin American countries undertook countrywide surveys or surveillance programs to determine their primary and acquired drug resistance prevalence rates. These studies followed the WHO/International Union Against Tuberculosis and Lung Diseases (IUATLD) guidelines. Percentages of not previously treated patients with tuberculosis due to MDR strains ranged from null or very small (Uruguay, Cuba, Chile) to 4% or higher (Dominican Republic, Argentina). In Argentina, a remarkable correlation between MDR tuberculosis, AIDS and the assistance in urban reference hospitals for infections diseases was observed. Coincidentally with the survey, nosocomial spread of HIV-related MDR tuberculosis occurred in two of these hospitals situated in Buenos Aires and Rosario. But, at the same time, an alarming emergence of MDR was evidenced among non HIV-infected patients with history of previous antituberculosis treatment. Directly observed treatment (DOT) should be increasingly applied, and drug supply guaranteed. Treatment as well as microscopy services for diagnosis and follow up of patients, should be decentralized from the big specialized hospitals in urban areas to the peripheral health centers, in order to make easier for the patients to attend regularly and receive their medications. These strategies will contribute to increase cure rates and to reduce the tuberculosis transmission.
Medicina 02/1998; 58(2):202-8. · 0.47 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: A steep upsurge of human immunodeficiency virus (HIV)-associated multidrug-resistant tuberculosis (MDR-TB) was recently observed at a referral treatment center in Buenos Aires City. Between January 1994 and June 1995, TB isolates resistant to at least five drugs were recovered from 101 of 272 HIV-infected inpatients. Highly resistant isolates from 77 patients underwent restriction fragment length polymorphism study with IS6110. After cross-contamination was eliminated, a single TB strain was found to have caused disease in 68 patients with a history of on-site exposure. The frequency of smear-positive pulmonary disease was higher among these patients than among non-MDR-TB HIV-infected patients (50/68 vs. 60/148, P < .001), and the 1-year survival was dramatically reduced (5/68 vs. 92/148). The strain involved in the outbreak was traced back to patients hospitalized in 1992. Institutional infection control policies were and may still be inadequate to contain the spread of TB among immunodepressed subjects, as is the case in other large urban hospitals in Argentina.
The Journal of Infectious Diseases 09/1997; 176(3):637-42. · 6.41 Impact Factor
-
Medicina 02/1996; 56(1):102-4. · 0.47 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Multidrug-resistant tuberculosis has emerged over the last two years at Carrasco Hospital, located in Rosario city. Nosocomial transmission among 7 AIDS patients admitted into the same ward between June and December/94 was supported by temporal clustering of cases, matching drug susceptibility, and identical IS6110 fingerprints. Among 8 non-HIV chronic cases without evidence of reciprocal contact outside the hospital, two additional clusters of 2 and 4 cases, respectively, were identified. The latter was found to be generated by a strain genetically related to the one that infected AIDS patients. It is hypothesized that an ancestor strain, common to both, might have been brought into the hospital long before the outbreak was first suspected.
Medicina 02/1996; 56(1):48-50. · 0.47 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The recent recovery of Mycobacterium paratuberculosis from tissues of patients with Crohn's disease has highlighted the possible etiologic role of this microorganism in the disease. However, the immunological evidence generated by various groups supporting this hypothesis is as yet inconclusive. A specific antibody response might be masked in these patients by the wide antigenic homologies prevailing within the genus Mycobacterium. The present study was undertaken with the purpose of exploring the humoral response to M. paratuberculosis in patients with Crohn's disease, by means of a cross-absorption procedure recently proposed for unveiling the presence of specific antibodies in bovine paratuberculosis. Antibodies IgG to M. paratuberculosis were investigated by enzyme-linked immunosorbent assay in 90 serum samples from 17 patients with Crohn's disease, 23 patients with ulcerative colitis an 14 with other bowel diseases. Samples from 86 subjects without bowel disease (healthy individuals and patients with tuberculosis, mycobacterioses and fungal diseases) were also included as controls. The specificity of these antibodies was explored by the absorption of sera with an ubiquitous Mycobacterium (M. phlei). The results were compared to those obtained by similar ELISA tests employing M. avium or M. tuberculosis as antigens. A faint humoral response to M. paratuberculosis and M. tuberculosis was detected in patients with Crohn's disease. Cross-absorption with M. phlei did not disclose a specific response nor was an increase in antibody levels detected in patients studied periodically. Sera from patients with ulcerative colitis and other bowel diseases also showed a slight reaction to mycobacteria.(ABSTRACT TRUNCATED AT 250 WORDS)
Medicina 02/1994; 54(2):97-102. · 0.47 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The effect of the human immunodeficiency virus (HIV) on mycobacterial antibody production was investigated. Using an enzyme-linked immunosorbent assay (ELISA) for detecting IgG against Mycobacterium tuberculosis PPD, it was observed that individuals at risk of HIV infection show a pattern of humoral response to the tubercle bacillus similar to that previously found in the immunocompetent population not exposed to risk factors: 6 of 12 (50.0%) tuberculosis cases had elevated levels of antibodies to PPD and 27 of 30 (90.0%) asymptomatic individuals had antibody levels within the normal range. In an HIV-seropositive group without AIDS indicator diseases, 8 of 22 (36.4%) tuberculous patients had detectable mycobacterial antibodies whereas 156 of 164 (95.1%) non-tuberculous subjects did not. Among AIDS cases, only 1 of 20 (5.0%) patients with tuberculosis and none of 53 non-tuberculous subjects showed a positive result. The study evidenced an increasing humoral unresponsiveness to PPD in the progression of HIV infection to AIDS. Thus, a serodiagnostic method for detecting tuberculosis such as the ELISA here employed noticeably decreases its utility in the latency stage of the HIV infection, and it is practically useless in clinical AIDS.
Tubercle and Lung Disease 09/1992; 73(4):187-91.
-
[show abstract]
[hide abstract]
ABSTRACT: A number of glycolipids were evaluated in an ELISA test for their serodiagnostic usefulness in tuberculosis. One hundred and twelve (112) sera belonging to bacteriologically confirmed TB patients, patients with pathologies other than tuberculosis and healthy individuals were examined against several synthetic "mirror" pseudo cord factors (analogues of trehalose-6,6'-dimycolate or TDM) using natural cord factor and another recently described natural glycolipid (SL-IV) of Mycobacterium tuberculosis as control antigens. Analysis of the results shows that all synthetic "mirror" pseudo cord factors, except one with a short 8-carbon chain, were better recognized by the sera of tuberculosis patients than natural cord factor, with sensitivity and specificity values in the ELISA test similar to those reported for M. tuberculosis species-specific SL-IV. Of all antigens tested in this study, BDA. TDA, a bis(N,N-dioctadecylamide) of "trehalose dicarboxylic acid", [(alpha-D-glucopyranosyluronic acid) (alpha-D-glucopyranosiduranic acid)], showed the highest serodiagnostic discriminating power (93% sensitivity and specificity). We postulate that either these artificial molecules are cross-reactants of similarly structured native glycolipids of M. tuberculosis or that they bear closer resemblance to actual phagosome-lysosome-modified antigens than to native mycobacterial ones.
Research in Microbiology 03/1992; 143(2):217-23. · 2.76 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: To assess the usefulness of enzyme immunoassay as a rapid method of diagnosing tuberculosis, a study was conducted of 687 serum samples from 271 children and 416 adults. With 55 sera from nontuberculous children as controls, the specificity was 0.98, and with 137 controls from the adult population, 0.93. Prior vaccination with BCG did not influence the level of detectable anti-PPD antibody. The results were similar in healthy PPD-positive and negative adults. The test differentiated mycoses and nontuberculous mycobacterioses from tuberculosis. The sensitivity rates in 49 children and 200 adults diagnosed with bacteriologically confirmed pulmonary and extrapulmonary tuberculosis were calculated at 0.51 and 0.69, respectively. In those tuberculosis cases not bacteriologically confirmed or at other sites, the test was positive in 28.1% of 114 children and in 48.6% of 35 adults. The cost, speed, and availability of reagents for this test were comparable to those for direct microscopic examination. Both methods were positive for 49% of the tuberculosis cases confirmed by culture, and a total of 84% of those cases were found positive using one method or the other. It is concluded that enzyme immunoassay can be especially useful in the rapid diagnosis of nonbacilliferous pulmonary, extrapulmonary, and childhood tuberculosis.
Boletín de la Oficina Sanitaria Panamericana. Pan American Sanitary Bureau 07/1991; 110(6):461-70.
-
[show abstract]
[hide abstract]
ABSTRACT: A sandwich ELISA for the detection of gamma interferon showed higher sensitivity and specificity than an indirect ELISA for mycobacterial antibodies in the diagnosis of bovine tuberculosis. Circumstantial evidence of an inverse relationship between cellular and humoral immune responses to Mycobacterium bovis was found in cattle with natural infection.
Research in Veterinary Science 06/1991; 50(3):365-7. · 1.65 Impact Factor
-
Medicina 02/1991; 51(1):77-9. · 0.47 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The sensitivity and specificity of an ELISA for the detection of bovine IgG anti-Mycobacterium bovis antibodies were 73.6% and 94.1%, respectively, as determined in 53 bacteriologically confirmed tuberculous cattle and 101 healthy cattle from a tuberculosis-free area. In addition, the results of ELISA and tuberculin tests in 149 cattle were compared with those of subsequent necropsy studies. Both tests failed to detect 2 animals with tuberculous lesions and positive culture; 3/12 cattle with M. bovis isolation and no lesions, and 2/7 with atypical mycobacterial infection reacted to tuberculin, but none had antibodies; in 128 cattle with neither lesions nor mycobacterial isolation, 6 were tuberculin reactors and 7 others had antibodies. Negative results were obtained by ELISA in 21/22 paratuberculous cattle. Antibodies were not detected in 88.9% to 96.4% of 697 cattle from two tuberculin negative herds of an endemic area. In a herd with proved M. bovis infection, distribution of seropositive animals in tuberculin and non-tuberculin reactors was similar. Antibody responses to cutaneous tuberculin stimuli were observed in 4 experimentally infected cattle, but only in 2/10 healthy controls after repeated PPD stimuli. Nine controls which had either received a single tuberculin dose or none showed no increase in antibody levels. The low sensitivity of this ELISA limits its usefulness as a diagnostic tool for bovine tuberculosis eradication campaigns. However, it could be helpful in epidemiological surveillance if its efficiency to identify infected herds is demonstrated.
Zentralblatt für Veterinärmedizin. Reihe B. Journal of veterinary medicine. Series B 03/1990; 37(1):19-27.
-
[show abstract]
[hide abstract]
ABSTRACT: An enzyme-linked immunosorbent assay for detecting antibodies to purified protein derivative was evaluated as a rapid method for serodiagnosis of childhood tuberculosis. Its specificity for IgG antibodies was 0.98 as determined in 55 sera from nontuberculous children who showed no significant effect of previous Bacillus Calmette-Guérin vaccination on the production of specific antibodies. Results were negative in 29 of 33 (87.9%) tuberculin-positive children and in 18 of 20 (90.0%) contacts, none of whom had evidence of tuberculosis. The sensitivity of this test was 0.51 as determined in 49 sera from bacteriologically confirmed cases; 17 of 27 smear positive cases and 8 of 22 children with positive cultures were detected. Results were positive in 32 of 114 (28.1%) patients with a diagnosis of tuberculosis not confirmed by microbiology. Consequently whereas a negative result does not rule out tuberculosis, a positive result is a strong indication of the disease. The IgM antibody determination yielded much less discriminative results.
The Pediatric Infectious Disease Journal 12/1989; 8(11):763-7. · 3.58 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The ELISA has been extensively evaluated as a serodiagnostic method for tuberculosis. However, there is scarce information about its application to cases that cannot be diagnosed by microscopic examination: those with closed lesions or undergoing early stages of the disease. Since a reliable serological test might substantially contribute to their prompt detection, the objective of the present study was to determine the diagnostic value of an ELISA applied to adult smear-negative cases of tuberculosis. Sera from 235 patients with active tuberculosis--176 pulmonary and 59 extrapulmonary cases--and 181 control subjects were tested for IgG antibodies to PPD by ELISA. Eleven cases of non tuberculous mycobacterial (MOTT) disease and 33 cases of mycosis were also included in this group. With the adopted cut-off value, 73.9% (105/142) of smear positive and 52.7% (49/93) of smear negative tuberculosis cases, were correctly classified. Particularly in the latter, the test was positive in 55.2% (32/58) of patients with positive cultures for Mycobacterium tuberculosis and in 48.6% (17/35) of patients diagnosed by clinical, radiological and or histopathological findings. No antibody activity was demonstrated in 92.7% of sera from the control population which included 92 healthy volunteers, 32 non tuberculous diseased subjects and 13 household contacts of smear-positive cases. Among those control subjects who were skin tested, ELISA results were not related to the tuberculin reactivity: 93.7% (30/32) of tuberculin negative and 95.2% (40/42) of tuberculin positive healthy individuals had no detectable antibodies.(ABSTRACT TRUNCATED AT 250 WORDS)
Medicina 02/1989; 49(6):561-6. · 0.47 Impact Factor
-
Bulletin of the Pan American Health Organization 02/1988; 22(3):269-80.
-
[show abstract]
[hide abstract]
ABSTRACT: Fifteen laboratories from six regions of the country participated in the present trial covering the three years period 1982-84 during which they obtained 13544 mycobacterial cultures from 7672 patients. Of the total isolates, 437 (3.22%) were nontuberculous mycobacteria and 49 (0.36%) Mycobacterium bovis. The remaining cultures were identified as M. tuberculosis, which was the infectious agent in 7609 cases (99.17%). A further 36 cases (0.47%) were due to M. bovis and the remaining 27 to nontuberculous mycobacteria (0.35%). This last group included 26 cases due to M. avium - intracellulare - scrofulaceum complex (MAIS) and one due to M. chelonae. All these were cases in adults with pulmonary lesions except for a lymphadenitis diagnosed in a child. According to these results, the frequency of bovine tuberculosis is low in Argentina. Most of these cases were found in rural and slaughterhouse workers. Non tuberculous mycobacterioses are also of low frequency in the country.
Tropical and geographical medicine 08/1987; 39(3):222-7.
-
Zentralblatt für Veterinärmedizin. Reihe B. Journal of veterinary medicine. Series B 04/1987; 34(2):119-25.
-
Medicina 02/1986; 46(5):636-8. · 0.47 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The in vitro activity of six antituberculous agents and 13 nonspecific antibiotics and sulfonamides against 72 isolates of different atypical mycobacterial strains obtained in Argentina and Uruguay, was evaluated by the proportions susceptibility method on Löwenstein-Jensen medium. A non-homogeneous response was observed in strains of the same species. The use of nonspecific antibiotics and sulfonamides in the treatment of mycobacterioses cases should therefore be decided according to the results of susceptibility testing on the strain isolated from the patient. The method used here could be easily adopted by laboratories where the proportions method is currently employed to test the susceptibility of M. tuberculosis to specific drugs.
Zentralblatt für Bakteriologie, Mikrobiologie, und Hygiene. Series A, Medical microbiology, infectious diseases, virology, parasitology 11/1985; 260(2):247-53.
