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[A simple enzyme test for proving the resistance to isoniazid of Mycobacterium tuberculosis strains].

Resistance to isoniazid is the consequence of mutation in some genes or genomic sequences. One of them is katG gene, which encode catalase-peroxidase. The activity of this enzyme can be revealed by simultaneous test catalase-peroxidase. The aim of the study was to assess the correlation between the results of catalase-peroxidase test and isoniazid- resistance of M. tuberculosis strains, in order to estimate the utility of this test in current practice. Material and method. We assessed the results of catalase- peroxidase test of 2268 identified M. tuberculosis strains (19 isolated from silicotuberculosis patients, 198 isolated from industrial dust and chemical polluted city patients, and 2051 strains isolated from other pulmonary tuberculosis patients), with known pattern of susceptibility/resistance. Six hundred twenty four strains (27.5%) had been INH mono-resistant or INH resistant associated with other resistance. It resulted a significant correlation between INH resistance and the negative result of catalase-peroxidase test (p <0.001). More than that, the negative result of peroxidase test was obtained in 63.5% of INH resistant strains. This simple and easy to do test, used for the strains after growth in primary culture, can he helpful in order to confirm the suspicion of INH resistance.

Antitubercular Agents↗
PubMed1991

Bromoform.

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Animals↗
PubMed1981

Co-trimoxazole sensitivity testing: comparison of separate and combined disk agar diffusion techniques.

Five hundred and seven strains of bacteria isolated from the urine of patients with significant bacteriuria (more than 10(8) colony-forming units per litre) were tested for sensitivity to co-trimoxazole by the agar diffusion technique. Each organism was tested with a combined disk containing trimethoprim and sulphamethoxazole in a primary sensitivity test and, at a standardised inoculum, with both a combined disk and separate disks of trimethoprim and sulphamethoxazole. The results show that combined disk testing does not always indicate the sensitivity patterns of the organisms being tested.

Agar↗
PubMed1994

[Bacteriological diagnosis of tuberculosis].

Microscopic examination and culture are still today essential elements of the bacteriological diagnosis of tuberculosis. Microscopic examination of a Ziehl fuchsin or auramine stained specimen allows detection of most strains in less than an hour. Culture on Löwenstein-Jensen medium is more sensitive than the microscopic examination and is required for identification and to measure sensitivity to antibiotics. Mycobacterium colonies, generally the causal agent in tuberculosis, usually grow within 28 days and are easily recognized by their "cauliflower" aspect. The niacin test is used for formal identification. Currently, radiometric respirometry allows detection of M. tuberculosis growth and provides antibiotic sensitivity results more rapidly, usually within 10 days. Use of this technique is however limited because the culture medium contains radioactive carbon. Genetic probes are on the other hand quite easy to use and allow identification of cultured bacteria in only a few hours. After polymerization chain reaction gene amplification, M. tuberculosis strains can be detected directly in the specimen within 2 or 3 hours, but in practice, this method has not become a routine laboratory technique, particularly due to lack of sufficient specificity and sensitivity. No other serologic tests are currently reliable enough for the diagnosis of tuberculosis. For cases with low-count specimens, there still is no reliable "on-the-spot" diagnostic test.

Bacteriological Techniques↗
PubMed1985

Rapid methods for counting mycobacteria--comparison of methods for extraction of mycobacterial adenosine triphosphate (ATP) determined by firefly luciferase assay.

A comparison of 5 different methods of extraction of adenosine 5'-triphosphate (ATP) from mycobacterial cells was carried out using Mycobacterium bovis, BCG as a model. ATP was measured using the luciferin-luciferase bioluminescence reaction. Boiling buffer extraction was the best method. The amount of ATP extracted correlated with the number of colony forming units over a wide range of count. Although great sensitivity in terms of number of bacilli detectable was not achieved the method was rapid and appears suitable for drug sensitivity testing of tubercle bacilli.

Adenosine Triphosphate↗