[Evaluation of a multiplex PCR assay to differentiate mycobacteria of the Mycobacterium tuberculosis complex in a reference laboratory].
Mycobacteria that cause tuberculosis in animals and humans belong to the Mycobacterium tuberculosis complex. Techniques for conventional diagnosis are time-consuming and do not differentiate between different strains belonging to the M. tuberculosis complex. The aim of this study was to evaluate a multiplex PCR assay applicable to mycobacteria in culture with the capacity to differentiate different strains belonging to the M. tuberculosis complex in a reference laboratory. Primers based on genomics regions of difference (RD) consisting in DNA segments that are present in M. tuberculosis, but differentially deleted in several members of M. tuberculosis complex were used in a PCR assay. The test was applied to 86 clinical isolates of mycobacteria. The pattern of amplification allowed differentiating between M. tuberculosis, M. bovis and M. bovis BCG in a single PCR reaction. This PCR multiplex assay may be used in a Reference Laboratory of Tuberculosis Diagnosis as a complementary test to differentiate mycobacteria strains belonging to the M. tuberculosis complex. This test significantly reduces the time period between culture and strain identification, and thus for could favor the adoption of better strain specific antimycobacterial regimens as well as identification of zoonotic transmission of M. bovis to humans.
Arráiz R
Nailet
N
Sección de Biología Molecular y Escuela de Bioanálisis, Centro de Investigaciones Endocrino-Metabólicas Dr. Félix Gómez, Facultad de Medicina, Universidad del Zulia, Maracaibo, Venezuela. narraiz@cantv.net
Romay B
Zolay
Z
Faría M
Nelba
N
D015373:Bacterial Typing Techniques; D001483:Base Sequence; D004269:DNA, Bacterial / Q000032:analysis; D016680:Genome, Bacterial; D006801:Humans; D008969:Molecular Sequence Data; D009163:Mycobacterium bovis / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D009169:Mycobacterium tuberculosis / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D009841:Oligonucleotides / Q000032:analysis; D016133:Polymerase Chain Reaction / Q000379:methods*; D014376:Tuberculosis / Q000175:diagnosis / Q000382:microbiology*
D004269:DNA, Bacterial; D009841:Oligonucleotides