Nucleotide sequences which hybridize specifically with bacterial

Details Nucleotide sequences which hybridize specifically with bacterial Nucleotide sequences which hybridize specifically with bacterial

1994-05-02

1997-07-22

Arthur, Lisa B.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

4353201, 536 2432, 536 2433, 935 77, 935 78, C12Q 168, C12N 1563, C07H 2104

Patent

active

056502729

DESCRIPTION:

BRIEF SUMMARY
BACKGROUND OF THE INVENTION

(i) Field of the Invention
The present invention relates to a nucleic sequence specific for the Mycobacterium avium-intracellulare complex, as well as particular fragments of this sequence, which are capable of acting as nucleic probes for the detection of a nucleic acid specific for strains belonging to the Mycobacterium avium-intracellulare complex or as nucleic primers in the amplification of DNA or RNA obtained from strains of the Mycobacterium avium-intracellulare complex in a biological sample.
The invention also relates to a method of detection of strains belonging to the Mycobacterium avium-intracellulare complex in a biological sample using nucleic acid fragments, as well as a kit for implementing this method.
(ii) Description of Related Art
The number of infections caused by opportunistic mycobacteria is increasing, partly because they are frequent in AIDS patients. According to bacteriological studies carried out over the past few years, the most important opportunistic bacteria are: M. kansasii, M. xenopi, M. simiae and especially mycobacteria belonging to the complex: M. avium-intracellulare (MAI complex). However, geographical variations exist in the incidence of the infection by these various opportunistic mycobacteria.
In a recent study carried out by the Laboratory of Bacteriology and Virology of the Pitie-Salpetriere hospital group directed by Professor J. Grosset, it was shown that among the 709 AIDS patients whose blood was cultured (2460 samples) 63 (8.9%) had positive haemocultures with mycobacteria. On the other hand, 84.1% of the positive haemocultures contained M. avium-intracellulare and 11.1% M. tuberculosis.
M. avium and M. intracellulare are two phenotypically related species indiscernible by conventional identification, and are therefore grouped into an MAI complex. MAI complex for the purposes of the present invention is understood to mean the strains as defined by SAITO et al. (J. Clin. Microbiol., August 1990, 28, p. 1694-1697). This complex brings together human and animal pathogens. In man non-infected by the HIV viruses, the infections are especially located in the lungs in adults and in the ganglia in children. MAI infections are diagnosed in 50 to 90% of AIDS patients. The locations are, in this case, preferably in the digestive system and are disseminated. MAI is also a pathogen which is important in veterinary medicine, affecting poultry, bovine, caprine and ovine species and the like. The animal pathogens, M. paratuberculosis and M. silvaticum, were recently introduced into the nomenclature as sub-species of M. avium (Thorel et al., Int. J. Syst. Bacteriol., 1990, 40, 254-260). It should also be noted that strains of M. paratuberculosis were isolated from patients suffering from Crohn's disease.
Using serology, 28 serotypes are recognised within MAI. Currently, serotypes 1 to 6, 8 to 11 and 21 are classified as M. avium and serotypes 7, 12 to 20 and 25 as M. intracellulare. An uncertainty remains as to the taxonomic position of serotypes 22 to 24 and 26 to 28. In man non-infected with the HIV viruses, the distribution of the serotypes is highly dispersed whereas in AIDS patients, only serotypes 1, 4, 8 and 9 are predominant (data of the National Centre of Reference for Mycobacteria). This distribution is similar to that observed in animals where serotypes 1, 2, 3, 4, 8 are preferably found. The French epidemiological situation for the M. avium strains isolated from AIDS patients is different from that observed in the United States or in Sweden. While in France serotype 8 is predominant, serotypes 4 or 6 are the ones found in these other two countries.
The serotypes (or serovars) mentioned in the present patent are as defined by P. J. Brennan et al. (Identification of A typical Mycobacteria by Thin-Layer Chromatography of Their Surface Antigens, Journal of Clinical Microbiology, Oct. 1978, p. 374-379.
Opportunistic mycobacteria pose numerous challenges to the microbiologist, the immunologist and the clinician. Indeed, unlike M. tuberculos

REFERENCES:
patent: 5314801 (1994-05-01), Nycz et al.
patent: 5422242 (1995-06-01), Young
Baess et al. (1983) Path. Microbiol. Immunol. Scand. 91:201-203.
Saito et al. (1990, Aug.) J. Clin. Micro. 28(8):1694-1697.
Thierry et al. (1993) J. Clin. Micro. 31(5):1048-1054.
Journal of Clinical Microbiology, "Rapid Identification Using a Specific DNA Probe of Mycobacterium avium Complex from Patients with Acquired Immunodeficienty Syndrome", Kiehn, et al.; vol. 25, No. 8; Aug. 1987, pp. 1551-1552.
Molecular Biology, "Detection and Identification of Mycobacteria by Amplification of Mycobacterial DNA", Hance et al.; vol. 3, No. 7, 1989 pp. 843-849.

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