title: Validation of a real-time PCR assay for the molecular identification of Mycobacterium tuberculosis.
abstract: Mycobacterium tuberculosis is the major cause of tuberculosis in humans. This bacillus gained prominence with the occurrence of HIV, presenting itself as an important opportunistic infection associated with acquired immunodeficiency syndrome (AIDS). The current study aimed to develop a real-time PCR using Eva Green technology for molecular identification of M. tuberculosis isolates. The primers were designed to Rv1510 gene. Ninety nine samples of M. tuberculosis and sixty samples of M. bovis were tested and no sample of the bovine bacillus was detected by the qPCR. Statistical tests showed no difference between the qPCR and biochemical tests used to identify the Mycobacterium tuberculosis. The correlation between tests was perfect with Kappa index of 1.0 (p < 0.001, CI = 0.84 - 1.0). The diagnostic sensitivity and specificity were 100% (CI = 95.94% - 100%) and 100% (CI = 93.98% - 100%). This qPCR was developed with the goal of diagnosing the bacillus M. tuberculosis in samples of bacterial suspension. TB reference laboratories (health and agriculture sectors), public health programs and epidemiological studies probably may benefit from such method.
authors.affiliation: Laboratório Nacional Agropecuário de Minas Gerais Pedro LeopoldoMG Brazil Laboratório Nacional Agropecuário de Minas Gerais, Pedro Leopoldo, MG, Brazil.
authors.forename: Mariana L
authors.initials: ML
authors.lastname: Sales
authors.affiliation: Laboratório Nacional Agropecuário de Minas Gerais Pedro LeopoldoMG Brazil Laboratório Nacional Agropecuário de Minas Gerais, Pedro Leopoldo, MG, Brazil.
authors.forename: Antônio Augusto
authors.initials: AA
authors.lastname: Fonseca Júnior
authors.affiliation: Laboratório Nacional Agropecuário de Minas Gerais Pedro LeopoldoMG Brazil Laboratório Nacional Agropecuário de Minas Gerais, Pedro Leopoldo, MG, Brazil.
authors.forename: Lívia
authors.initials: L
authors.lastname: Orzil
authors.affiliation: Laboratório Nacional Agropecuário de Minas Gerais Pedro LeopoldoMG Brazil Laboratório Nacional Agropecuário de Minas Gerais, Pedro Leopoldo, MG, Brazil.
authors.forename: Andrea Padilha
authors.initials: AP
authors.lastname: Alencar
authors.affiliation: EMBRAPA Gado de Leite Juiz de ForaMG Brazil EMBRAPA Gado de Leite, Juiz de Fora, MG, Brazil.
authors.forename: Marcio Roberto
authors.initials: MR
authors.lastname: Silva
authors.affiliation: Laboratório Nacional Agropecuário de Minas Gerais Pedro LeopoldoMG Brazil Laboratório Nacional Agropecuário de Minas Gerais, Pedro Leopoldo, MG, Brazil.
authors.forename: Marina Azevedo
authors.initials: MA
authors.lastname: Issa
authors.affiliation: Laboratório Nacional Agropecuário de Minas Gerais Pedro LeopoldoMG Brazil Laboratório Nacional Agropecuário de Minas Gerais, Pedro Leopoldo, MG, Brazil.
authors.forename: Paulo Martins
authors.initials: PM
authors.lastname: Soares Filho
authors.affiliation: Escola de Veterinária Universidade Federal de Minas Gerais Belo HorizonteMG Brazil Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil.
authors.forename: Andrey Pereira
authors.initials: AP
authors.lastname: Lage
authors.affiliation: Escola de Veterinária Universidade Federal de Minas Gerais Belo HorizonteMG Brazil Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil.
authors.forename: Marcos Bryan
authors.initials: MB
authors.lastname: Heinemann
mesh_terms: D017931:DNA Primers / Q000235:genetics; D006801:Humans; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014376:Tuberculosis / Q000175:diagnosis*
chemical_list: D017931:DNA Primers
keywords: Mycobacterium tuberculosis; diagnosis; real time PCR