The hsp65 gene patterns of less common Mycobacterium and Nocardia spp. by polymerase chain reaction-restriction fragment length polymorphism analysis with capillary electrophoresis

Po-Ling Chang; Wen-Shyang Hsieh; Chia-Lien Chiang; Marion J Tuohy; Gerri S Hall; Gary W Procop; Huan-Tsung Chang; Hsin-Tsung Ho

doi:10.1016/j.diagmicrobio.2007.02.004

The hsp65 gene patterns of less common Mycobacterium and Nocardia spp. by polymerase chain reaction-restriction fragment length polymorphism analysis with capillary electrophoresis

Diagn Microbiol Infect Dis. 2007 Jul;58(3):315-23. doi: 10.1016/j.diagmicrobio.2007.02.004. Epub 2007 Mar 26.

Authors

Po-Ling Chang¹, Wen-Shyang Hsieh, Chia-Lien Chiang, Marion J Tuohy, Gerri S Hall, Gary W Procop, Huan-Tsung Chang, Hsin-Tsung Ho

Affiliation

¹ Department of Chemistry, National Taiwan University, Taipei, Taiwan.

PMID: 17382507
DOI: 10.1016/j.diagmicrobio.2007.02.004

Abstract

To rapidly identify Mycobacterium and Nocardia spp. without costly probes, we had implemented capillary electrophoresis (CE) in polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis to analyze their 65-kDa heat shock protein (hsp65) gene. The PCR-RFLP analysis with CE (PRACE) involved only one restriction enzyme, HaeIII, and a single electrophoretic separation less than 10 min. Full-range (10-200 bp) RFLP patterns of 12 less common Mycobacterium and 7 Nocardia spp. were investigated. A good agreement was observed between the sizes of restriction fragments resolved by CE and the real sizes deduced from sequence analysis. Including hsp65 gene patterns of 12 Mycobacterium spp. published earlier, differentiation was distinct among 24 Mycobacterium and 7 Nocardia spp. Some closely related species exhibiting similar biochemical characteristics could be well discriminated by an extra HaeIII digestion site. Thus, PRACE offers a nonprobe alternative for rapid identification of various cultured Mycobacterium and Nocardia to the species level.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bacterial Proteins / genetics*
Chaperonin 60
Chaperonins / genetics*
DNA, Bacterial / genetics
Deoxyribonucleases, Type II Site-Specific
Electrophoresis, Capillary / methods*
Mycobacterium / classification*
Mycobacterium / genetics
Nocardia / classification*
Nocardia / genetics
Polymerase Chain Reaction
Polymorphism, Restriction Fragment Length*

Substances

Bacterial Proteins
Chaperonin 60
DNA, Bacterial
heat-shock protein 65, Mycobacterium
Deoxyribonucleases, Type II Site-Specific
GGCC-specific type II deoxyribonucleases
Chaperonins