1887

Abstract

A repetitive sequence specific to was isolated from a gt11 library of by DNA–DNA hybridization using genomic DNA of as probe followed by subtractive hybridization with a cocktail of other mycobacterial DNA. This led to identification of CD192, a 1291 bp fragment of containing repetitive sequences, which produced positive hybridization signals with DNA within 30 min. Nucleotide sequencing revealed the presence of several direct and inverted repeats within the 1291 bp fragment that belonged to a PPE family gene (Rv0355) of . The use of CD192 as a DNA probe for the identification of in culture and clinical samples was investigated. The 1291 bp sequence was present in , and BCG, but was not present in many of the other mycobacterial strains tested, including H37Ra. More than 300 clinical isolates of were probed with CD192, and the presence of the 1291 bp sequence was observed in all the clinical strains, including those lacking IS. The sequence displayed RFLP among the clinical isolates. A PCR assay was developed which detected with 100 % specificity from specimens of sputum, cerebrospinal fluid and pleural effusion from clinically diagnosed cases of tuberculosis.

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2006-08-01
2019-11-18
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