Rapid amplification-based subtyping of mycobacterium tuberculosi

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid

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435 912, 435 9152, 536 2432, C12Q 168

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active

056521065

ABSTRACT:
The present invention provides methods of detecting or distinguishing the DNA of an individual strain of Mycobacterium tuberculosis utilizing the polymerase chain reaction (PCR). Reproducible, unique patterns can be produced allowing the identification of unknown M. tuberculosis DNA by performing this reaction and comparing the pattern produced to the known reproducible, unique patterns. The invention further provides a kit useful to detect or distinguish the DNA of an individual strain of M. tuberculosis in a sample, comprising specific primers for use in PCR. The present invention also provides a method of determining the presence of a multidrug-resistant M. tuberculosis by detecting the presence of a specific arrangement of genomic DNA. Such detection can be done using PCR or a ligase chain reaction (LCR). The present invention provides nucleic acid sequences useful in detecting multidrug-resistant M. tuberculosis.

REFERENCES:
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Edlin, et al., "An Outbreak of Multidrug-Resistant Tuberculosis Among Hospitalized Patients With the Acquired Immunodeficiency Syndrome," The New England Journal of Medicine 326(23):1514-1521 (Jun. 4, 1992).
Hermans, et al., "Characterization of a Major Polymorphic Tandem Repeat in Mycobacterium tuberculosis and its Potential Use in the Epidemiology of Mycobacterium kansasii and Mycobacterium gordonae," Journal of Bacteriology 174(12):4157-4165 (Jun. 1992).
Cousins, et al., "Use of Polymerase Chain Reaction for Rapid Diagnosis of Tuberculosis," Journal of Clinical Microbiology 30(1):255-258 (Jan. 1992).
Otal, et al., "Restriction Fragment Length Polymorphism Analysis Using IS110 as an Epidemiological Marker in Tuberculosis," Journal of Clinical Microbiology 29(6):1252-1254 (Jun. 1991).
Cave, et al., "IS6110: Conservation of Sequence in the Mycobacterium tuberculosis Complex and its Utilization in DNA Fingerprinting," Molecular and Cellular Probes 5:73-80 (1991).
Versalovic, et al., "Distribution of Repetitive DNA Sequences in Eubacteria and Application to Fingerprinting of Bacterial Genomes," Nucleic Acids Research 19(24):6823-6831 (1991).
Thomas M.Shinnick, "The 65-Kilodalton Antigen of Mycobacterium tuberculosis,"Journal of Bacteriology 169(3):1080-1088 (Mar. 1987).

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