Development of a 23S rRNA-based PCR assay for the detection of mycobacteria.
Indian J Biochem Biophys
;
1994 Aug; 31(4): 288-94
Article
in English
| IMSEAR
| ID: sea-27463
ABSTRACT
The partial nucleotide sequence of a recombinant plasmid containing the 23S rRNA gene of Mycobacterium tuberculosis was determined and an assay was developed for amplifying 23S rRNA gene sequences of mycobacteria. The PCR-based non-radioactive test enabled us to distinguish Mycobacterium from other closely related genera and was sensitive enough to detect 2 bacterial genome equivalents. The assay was extended to the detection of mycobacterial DNA in uncultured clinical specimens; 23S rRNA sequences were detected in thirty four of forty eight (70.8%) sputum and cerebrospinal fluid (CSF) specimens by the PCR assay, whereas direct smear examination and culture methods demonstrated a positivity rate of 29.2% and 16.7% respectively for the same specimens. A RNA-based PCR assay with a detection limit of 1 genome equivalent was also developed. These PCR assays should prove useful for the early and rapid detection of mycobacterial infection in uncultured clinical specimens.
Full text:
Available
Index:
IMSEAR (South-East Asia)
Main subject:
Humans
/
Molecular Sequence Data
/
RNA, Ribosomal, 23S
/
Base Sequence
/
Polymerase Chain Reaction
/
Sensitivity and Specificity
/
Mycobacterium tuberculosis
Type of study:
Diagnostic study
Language:
English
Journal:
Indian J Biochem Biophys
Year:
1994
Type:
Article
Similar
MEDLINE
...
LILACS
LIS