Purification and characterisation of isocitrate dehydrogenase and malate dehydrogenase from Mycobacterium tuberculosis and evaluation of their potential as suitable antigens for the serodiagnosis of tuberculosis.
METHODS
Enzymes from Mycobacterium tuberculosis are potent antigens and might thus be of interest in the serodiagnosis of tuberculosis.


OBJECTIVE
The purpose of the study was to purify and characterize the two enzymes isocitrate dehydrogenase (IDH) and malate dehydrogenase (MDH) from, M. tuberculosis and to evaluate their potential in the serodiagnosis of tuberculosis.


METHODS
The two enzymes were analysed for specificity by electrophoresis and then purified by means of affinity chromatography using reactive dyes and ion exchange chromatography. The two isolated enzyme fractions were analysed by ELISA, using antisera against related organisms. They were then tested as antigens in ELISA together with sera from tuberculous patients and controls.


RESULTS
The electrophoretical analyses showed that the two enzymes each differed markedly from the corresponding enzymes of other mycobacteria. The serological analyses, however, could not distinguish between either IDH or MDH from other mycobacteria, but organisms of other genera, such as Nocardia, gave much weaker responses. When IDH and MDH were tested with sera from tuberculous patients and controls the former gave clearly higher optical density values than the latter.


CONCLUSIONS
The enzymes/antigens IDH and MDH may be of value in developing a serological test for tuberculosis. The latter fraction seemed particularly capable of discriminating patients from controls.
Ohman
R
R
Department of Medical Microbiology and Immunology, Göteborg University, Sweden.
Ridell
M
M
D000818:Animals; D000942:Antigens, Bacterial; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D007521:Isocitrate Dehydrogenase / Q000276:immunology / Q000302:isolation & purification*; D008291:Malate Dehydrogenase / Q000276:immunology / Q000302:isolation & purification*; D009169:Mycobacterium tuberculosis / Q000201:enzymology* / Q000276:immunology; D011817:Rabbits; D014376:Tuberculosis / Q000175:diagnosis*
D000942:Antigens, Bacterial; D008291:Malate Dehydrogenase; D007521:Isocitrate Dehydrogenase