Evaluation of a Single-Tube Multiplex Real-Time PCR for Differentiation of Members of the Mycobacterium tuberculosis Complex in Clinical Specimens

• DOI: 10.1128/jcm.00467-11
• Publication Date: 2011-05-19T03:55:42Z
• Authors: Tanya A. Halse; Vincent E. Escuyer; Kimberlee A. Musser
• Keywords: Mycobacterium tuberculosis complex; Multiplex; Concordance; Nontuberculous Mycobacteria

Members of the Mycobacterium tuberculosis complex (MTBC) differ in virulence attributes, drug resistance patterns, and host preferences. The rapid differentiation these species to determine zoonotic or human sources disease direct treatment can benefit both public health patient management. Commercially available assays cannot differentiate species, published have not been evaluated directly on clinical specimens. A real-time PCR assay for M. tuberculosis, bovis, bovis BCG, africanum, microti, canettii was developed. presence absence regions difference (RD) between genomes members MTBC allowed design a single-tube five-plex species. This assesses RD1, RD4, RD9, RD12, region exterior RD9 which is present all members. To evaluate performance this assay, 192 specimens positive by were tested, resulting 94% correlation with identification results obtained cultured material. Additionally, 727 Bactec MGIT 960-positive cultures 97% concordance methods. an inexpensive (2.5-h) method performed closed-format system requiring minimal hands-on time that be implemented laboratory used