Identification and subspecific differentiation of Mycobacterium scrofulaceum by automated sequencing of a region of the gene (hsp65) encoding a 65-kilodalton heat shock protein

J Clin Microbiol. 1996 Dec;34(12):3151-9. doi: 10.1128/jcm.34.12.3151-3159.1996.

Abstract

Mycobacterium scrofulaceum is most commonly recovered from children with cervical lymphadenitis, although it also accounts for approximately 2% of the mycobacterial infections in AIDS patients. Species assignment of M. scrofulaceum isolated by conventional techniques can be difficult and time-consuming. To develop a strategy for rapid species assignment of these organisms, a 360-bp region of the gene (hsp65) encoding a 65-kDa heat shock protein in 37 isolates from diverse sources was sequenced. Eight hsp65 alleles were identified, and these sequences formed phylogenetic clusters and lineages largely distinct from other Mycobacterium species. There was incomplete correlation between serovar designation and hsp65 allele assignment. The hsp65 data correlated strongly with the results of sequence analysis of the gene coding for 16S rRNA. Automated DNA sequencing of a 360-bp region of the hsp65 gene provides a rapid and unambiguous method for species assignment of these acid-fast organisms for diagnostic purposes.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Alleles
  • Amino Acid Sequence
  • Bacterial Proteins*
  • Bacterial Typing Techniques
  • Base Sequence
  • Chaperonin 60
  • Chaperonins / genetics*
  • Child
  • DNA Primers / genetics
  • DNA, Ribosomal / genetics
  • Genes, Bacterial*
  • Humans
  • Lymphadenitis / diagnosis
  • Lymphadenitis / microbiology
  • Molecular Sequence Data
  • Mycobacterium Infections, Nontuberculous / diagnosis
  • Mycobacterium Infections, Nontuberculous / microbiology
  • Mycobacterium scrofulaceum / classification*
  • Mycobacterium scrofulaceum / genetics*
  • Mycobacterium scrofulaceum / isolation & purification
  • Phylogeny
  • RNA, Bacterial / genetics
  • RNA, Ribosomal, 16S / genetics
  • Sequence Analysis, DNA
  • Sequence Homology, Nucleic Acid
  • Serotyping
  • Species Specificity

Substances

  • Bacterial Proteins
  • Chaperonin 60
  • DNA Primers
  • DNA, Ribosomal
  • RNA, Bacterial
  • RNA, Ribosomal, 16S
  • heat-shock protein 65, Mycobacterium
  • Chaperonins

Associated data

  • GENBANK/U17955
  • GENBANK/X52924