Your browser doesn't support javascript.
loading
A two-tube combined TaqMan/SYBR Green assay to identify mycobacteria and detect single global lineage-defining polymorphisms in Mycobacterium tuberculosis.
Cheah, Eddy S G; Malkin, Joanne; Free, Robert C; Lee, Su-Min; Perera, Nelun; Woltmann, Gerrit; Patel, Hemu; Kimmitt, Patrick T; Smith, Rebecca J; Rajakumar, Kumar; Barer, Michael R.
Afiliación
  • Cheah ES; Department of Infection, Immunity, and Inflammation, The Medical School, University of Leicester, Leicester, United Kingdom.
J Mol Diagn ; 12(2): 250-6, 2010 Mar.
Article en En | MEDLINE | ID: mdl-20093392
We have developed a novel real-time PCR assay to identify and perform preliminary genotyping of mycobacteria in a manner tailored to our local service. Within a single thermocycler run, mycobacterial 16S rDNA and the Mycobacterium tuberculosis global lineage-defining RD750 polymorphism are targeted in separate reaction tubes, each of which includes both TaqMan and SYBR Green chemistries. The results of this 16S-RD assay differentiate M. tuberculosis complex (MTBC) from nontuberculous mycobacteria (NTM) and recognize whether or not MTBC isolates belong to the East African-Indian lineage, the single most frequently isolated global MTBC lineage in our service. If required, NTM amplicons may be sequenced to provide more specific identities. We report the technical performance of this assay on 88 mycobacteria-positive cultures and discuss its use in the initial management of mycobacterial infections. The 16S-RD assay correctly identified all 70 MTBC-positive cultures and 17 NTM-positive cultures while contemporaneously recognizing 26 MTBC isolates as within and 44 outside the East African-Indian lineage. In artificial samples, the combined assay also showed limited potential to detect mixed mycobacterial infections (MTBC/NTM) and tuberculosis infections involving more than one global MTBC lineage. The approach we have established can be readily tailored to targets of particular value for any mycobacterial diagnostic service, thereby optimizing the value of the results for local clinical and public health management of mycobacterial infections.
Asunto(s)
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Compuestos Orgánicos / Polimorfismo Genético / Reacción en Cadena de la Polimerasa / Técnicas de Tipificación Bacteriana / Polimerasa Taq / Colorantes Fluorescentes / Mycobacterium tuberculosis Tipo de estudio: Evaluation_studies Límite: Humans Idioma: En Revista: J Mol Diagn Asunto de la revista: BIOLOGIA MOLECULAR Año: 2010 Tipo del documento: Article País de afiliación: Reino Unido Pais de publicación: Estados Unidos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Compuestos Orgánicos / Polimorfismo Genético / Reacción en Cadena de la Polimerasa / Técnicas de Tipificación Bacteriana / Polimerasa Taq / Colorantes Fluorescentes / Mycobacterium tuberculosis Tipo de estudio: Evaluation_studies Límite: Humans Idioma: En Revista: J Mol Diagn Asunto de la revista: BIOLOGIA MOLECULAR Año: 2010 Tipo del documento: Article País de afiliación: Reino Unido Pais de publicación: Estados Unidos