Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1995-05-03
1999-10-05
Housel, James C.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
530350, 4242001, 4242031, 4242481, G01N 33554
Patent
active
059622407
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to Mycobacterium proteins, in particular those of M. bovis, having molecular weights between approximately 44.5 and 47.5 kD and the nucleotide sequences coding for these proteins.
The present invention also relates to those protein fractions obtained from cultures of Mycobacterium bovis, showing a specific immunological reactivity towards anti-tuberculosis antibodies.
It also relates to the use of these proteins and fractions for the detection and monitoring of tuberculosis and as vaccines.
Tuberculosis continues to be a public health problem throughout the world. The annual number of deaths directly related to tuberculosis is around 3 million and the number of new cases of tuberculosis is around 15 million. This number of deaths due to tuberculosis is high even for the developed countries; for example in France it is of the order of 1500 per year, a figure which is certainly underestimated by a factor of 2 or 3 if Roujeau's assessments of the differences between official figures and the results of systematic autopsies are taken into account. The recent increase in tuberculosis cases, or at least the leveling-off of the decrease in the frequency of this disease, must be considered in correlation with the development of the HIV/AIDS epidemic. In total tuberculosis remains the leading infectious disease in terms of frequency in France and the developed countries, but above all in the developing countries for which it constitutes the principal source of human loss related to a single disease.
At present, a definite diagnosis made by the demonstration of cultivatable bacilli in a sample taken from the patient is only obtained in less than half the cases of tuberculosis. Even for pulmonary tuberculosis, which represents 80 to 90% of the tuberculosis cases, and which is the form of the disease for which the detection of the bacilli is the easiest, the examination of expectorations is only positive for less than half the cases.
The development of the most sensitive techniques such as PCR (Polymerized Chain Reaction), always comes up against the necessity for obtaining a sample. Since women and children do not habitually spit, sampling for infants frequently requires specialized medical intervention (for example ganglionic biopsy or sampling by lumbar puncture of the cephalo-rachidian fluid).
In other respects, inhibitions of the PCR reaction itself exist, of a type such that a sample can be unusable by this technique because of the impossibility of controlling their origins.
Finally, the conventional bacteriological diagnosis, microscopic examination and culture, because of its sensitivity limits (at the best of the order of 10.sup.4 to 10.sup.5 bacilli in the sample) requires that there has already been a relatively substantial development of bacilli and thus of the disease.
The detection of specific antibodies directed against Mycobacterium tuberculosis should thus be of assistance in the diagnosis of the common forms of the disease for which the detection of the bacilli themselves is difficult or impossible.
Successive generations of research workers have attempted to perfect a serological diagnosis technique for tuberculosis. From Ardoing (C.r. hebd. Seanc. Acad. Sci. Paris; 1898: 126: 1398-1401) to Middlebrook and Dubos (J. Exp. Med. 1948, 88: 521-528), the preparations used for this diagnosis have been very little or not purified, the effort being directed above all towards an increase in sensitivity and not specificity. Recently again techniques tending to increase only the sensitivity have been suggested using a technique of either the ELISA or RIA type.
The more recent work of Daniel and Janicki (Microbiol. Rev. 1978, 42; 84-113) or Wiker et al. (Scand. J. Immunol. 1988, 27: 223-239) has shown the complexity of the mycobacterial antigens. Following this work, attempts have been made to define the principal antigens able to be used for diagnostic purposes (Chan et al., Am. Rev. Respir. Dis. 1990, 142: 385-390). A serological test has as a result been commercially available (A
REFERENCES:
Abou-Zeid et al. Infect. Immun. Dec. 1987. 55(12):3213-3214.
Miura et al. Infect Immun. Feb. 1983. 39:540-545.
Fifis et al. Infect Immun. Mar. 1991. 59(3):800-807.
DeBruyn et al. Infect. Immun. 1987. 55(1):242-252.
Marchal Gilles
Pescher Pascale
Romain Felix
Graser Jennifer
Housel James C.
Institut Pasteur
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