A highly sensitive quantitative real-time PCR assay based on the groEL gene of contemporary Thai strains of Orientia tsutsugamushi

Clin Microbiol Infect. 2009 May;15(5):488-95. doi: 10.1111/j.1469-0691.2008.02671.x. Epub 2009 Mar 11.

Abstract

Partial nucleotide sequences (459 bp) of the groEL gene (encoding the 60-kDa heat shock protein, HSP60) from 23 contemporary isolates of Orientia tsutsugamushi isolated from patients with acute scrub typhus in Thailand were compared with 16 reference strain sequences to evaluate the potential of groEL as a conserved and representative target for molecular diagnostics.. Overall nucleotide identity within all available O. tsutsugamushi isolates (n = 39) was 98.8% (range: 95.0-100), reflecting a high degree of conservation; nucleotide identities were 67.5% and 65.6%, respectively, when typhus and spotted fever group rickettsiae were included.. A highly sensitive and quantitative real-time PCR assay was designed and evaluated using 61 samples, including buffy coats from patients in Thailand and Laos. Reliable and accurate quantitation of bacterial loads allows further investigation of other diagnostic methods and may lead to an improved understanding of the pathophysiology of acute scrub typhus, an important but under-recognized disease.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chaperonin 60 / genetics*
  • Conserved Sequence
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • Humans
  • Laos
  • Molecular Sequence Data
  • Orientia tsutsugamushi / genetics*
  • Orientia tsutsugamushi / isolation & purification*
  • Phylogeny
  • Polymerase Chain Reaction / methods*
  • Scrub Typhus / diagnosis*
  • Scrub Typhus / microbiology
  • Sensitivity and Specificity
  • Sequence Analysis, DNA
  • Sequence Homology
  • Sequence Homology, Nucleic Acid
  • Thailand

Substances

  • Chaperonin 60
  • DNA, Bacterial

Associated data

  • GENBANK/EF551288
  • GENBANK/EF551289
  • GENBANK/EF551290
  • GENBANK/EF551291
  • GENBANK/EF551292
  • GENBANK/EF551293
  • GENBANK/EF551294
  • GENBANK/EF551295
  • GENBANK/EF551296
  • GENBANK/EF551297
  • GENBANK/EF551298
  • GENBANK/EF551299
  • GENBANK/EF551300
  • GENBANK/EF551301
  • GENBANK/EF551302
  • GENBANK/EF551303
  • GENBANK/EF551304
  • GENBANK/EF551305
  • GENBANK/EF551306
  • GENBANK/EF551307
  • GENBANK/EF551308
  • GENBANK/EF551309
  • GENBANK/EF551310