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

被引：18

作者：

Chang, Po-Ling

Hsieh, Wen-Shyang

Chiang, Chia-Lien

Tuohy, Marion J.

Hall, Gerri S.

Procop, Gary W.

Chang, Huan-Tsung ^{[1
]}

Ho, Hsin-Tsung

机构：

[1] Natl Taiwan Univ, Dept Chem, Taipei 10764, Taiwan

[2] Mackay Mem Hosp, Dept Lab Med, Taipei, Taiwan

[3] Cleveland Clin, Dept Clin Pathol, Clin Microbiol Sect, Cleveland, OH 44106 USA

[4] Natl Taitung Univ, Dept Nat Sci Educ, Taitung, Taiwan

[5] Mackay Med Nursing & Management Coll, Taipei, Taiwan

来源：

DIAGNOSTIC MICROBIOLOGY AND INFECTIOUS DISEASE | 2007年 / 58卷 / 03期

关键词：

restriction fragment length polymorphism; PCR-RFLP analysis; capillary electrophoresis; Mycobacteria other than tuberculosis; Nocardia;

D O I：

10.1016/j.diagmicrobio.2007.02.004

中图分类号：

R51 [传染病];

学科分类号：

100401 ;

摘要：

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. (c) 2007 Elsevier Inc. All rights reserved.

引用

页码：315 / 323

页数：9

共 44 条

[21] DIFFERENTIATION OF SLOWLY GROWING MYCOBACTERIUM SPECIES, INCLUDING MYCOBACTERIUM-TUBERCULOSIS, BY GENE AMPLIFICATION AND RESTRICTION-FRAGMENT-LENGTH-POLYMORPHISM ANALYSIS [J].