Identification of Mycobacterium tuberculosis complex by polymerase chain reaction of Exact Tandem Repeat-D fragment from mycobacterial cultures

C Coitinho; G Greif; C Robello; J van Ingen; C Rivas

doi:10.1016/j.ijmyco.2012.07.002

Identification of Mycobacterium tuberculosis complex by polymerase chain reaction of Exact Tandem Repeat-D fragment from mycobacterial cultures

Int J Mycobacteriol. 2012 Sep;1(3):146-8. doi: 10.1016/j.ijmyco.2012.07.002. Epub 2012 Aug 27.

Authors

C Coitinho¹, G Greif², C Robello³, J van Ingen⁴, C Rivas⁵

Affiliations

¹ Comisión Honoraria para la Lucha Antituberculosa y Enfermedades Prevalentes, Centro de Referencia Nacional para Micobacterias, Departamento de Laboratorio, 18 de Julio av. 2175 6p, CP 11200 Montevideo, Uruguay. Electronic address: ccoitinho@yahoo.com.
² Institut Pasteur de Montevideo, Mataojo 2020, CP 11400 Montevideo, Uruguay.
³ Institut Pasteur de Montevideo, Mataojo 2020, CP 11400 Montevideo, Uruguay; Depto. de Bioquímica, Facultad de Medicina, Gral. Flores 2125, CP 11800 Montevideo, Uruguay.
⁴ Department of Clinical Microbiology, Radboud University Nijmegen Medical Center, 6500 HC, P.O. Box 9102, Nijmegen, The Netherlands.
⁵ Comisión Honoraria para la Lucha Antituberculosa y Enfermedades Prevalentes, Centro de Referencia Nacional para Micobacterias, Departamento de Laboratorio, 18 de Julio av. 2175 6p, CP 11200 Montevideo, Uruguay.

PMID: 26787211
DOI: 10.1016/j.ijmyco.2012.07.002

Abstract

This study evaluated an in-house polymerase chain reaction (PCR) for rapid identification of the Mycobacterium tuberculosis complex (MTBC) using the MTBC-specific Exact Tandem Repeat D (ETR-D) as the amplification target. In a prospective study, 801 clinical isolates identified as MTBC and 15 nontuberculous mycobacteria were analyzed. Mycobacterial DNA was extracted from automated broth cultures or from egg-based media. The amplification of the ETR-D showed to a sensitivity of 99.6% and a specificity of 100% for the correct identification of MTBC; improved extractions protocols led to 100% sensitivity. The main utility of this technique is the simplicity, rapidity, low cost and accuracy.

Keywords: Diagnosis; ETR-D; Identification; M. tuberculosis complex.