Polymerase Chain Reaction Amplification of a Repetitive DNA Sequence Specific for Mycobacterium tuberculosis
Abstract
A segment of DNA repeated in the chromosome of Mycobacterium tuberculosis was sequenced and used as a target for amplification using polymerase chain reaction (PCR). The sequences of the primers (5' to 3') were CCTGCGAGCGTAGGCGTCGG and CTCGTCCAGCGCCGCTTCGG, and a temperature of 68 degrees C was used for annealing the primers in the reaction. Amplification produced a 123-base-pair fragment with an internal SalI site. The specific PCR product was obtained with input DNA from 11 different strains of M. tuberculosis and Mycobacterium bovis and one strain of Mycobacterium simiae. No product was detected with DNA from 28 strains of the Mycobacterium avium complex, Mycobacterium scrofulaceum, Mycobacterium kansasii, Mycobacterium fortuitum, Mycobacterium chelonei, and Mycobacterium gordonae. The PCR product was detected by gel electrophoresis after 30 cycles using 1 fg of input DNA. Amplification of this sequence may provide the basis for an assay to detect M. tuberculosis directly in clinical material.
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False-positive reactions are a major problem with PCR methods for detection of M. tuberculosis.
“False-positive reactions are a major problem with PCR methods for the detection of the M. tuberculosis (Eisenach et al., 1990; Friess et al., 1990; DelPortillo et al., 1991; Eisenach et al., 1991).”
From Application of PCR from the fine needle aspirates for the diagnosis of cervical tuberculous lymphadenitis. (1996) · cites this paper as Eisenach et al., 1990
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- Authors
- Eisenach, K. D., Donald Cave, M., Bates, J. H., & Crawford, J. T.
- Affiliations
- John L. McClellan Memorial Veterans Hospital (US)
- ORCID
- Eisenach, K. D.
- Venue
- Journal of Infectious Diseases
- Publisher
- Oxford University Press (OUP)
- Published
- 1990-05-01
- Volume
- 161
- Issue
- 5
- Pages
- 977-981
- ISSN
- 0022-1899 (ISSN-L)
- 1537-6613
- OpenAlex
- W1968649252
- Cited by
- 748
- References
- 0
- Citations by year
- 2026: 6 · 2025: 2 · 2024: 7 · 2023: 11 · 2022: 11 · 2021: 12 · 2020: 8 · 2019: 11 · 2018: 9 · 2017: 21 · 2016: 21 · 2015: 21 · 2014: 18 · 2013: 27 · 2012: 29
- FWCI
- 39.00
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- The Journal of Infectious Diseases (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Health Sciences › Medicine › Infectious Diseases › Tuberculosis Research and Epidemiology
- Health Sciences › Medicine › Epidemiology › Mycobacterium research and diagnosis
- Health Sciences › Medicine › Surgery › Infectious Diseases and Tuberculosis
- Keywords
- Mycobacterium tuberculosis, repetitive DNA, PCR detection, diagnostic assay, primer design, Mycobacterium bovis, Mycobacterium simiae, Mycobacterium avium complex
- MeSH terms
- Base Sequence, Cloning, Molecular, DNA, Bacterial, Electrophoresis, Agar Gel, Gene Amplification, Humans, Molecular Sequence Data, Mycobacterium tuberculosis, Nucleic Acid Hybridization, Repetitive Sequences, Nucleic Acid, Temperature, Polymerase Chain Reaction
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