• 作者：Laura Herrera-Leó ; n ; ^{lherrera@isciii.es} ; Rodolfo Pozuelo-Dí ; az ; Tamara Molina Moreno ; Azucena Valverde Cobacho ; Pilar Saiz Vega ; Marí ; a Soledad Jimé ; nez Pajares
• 关键词：Complejo Mycobacterium tuberculosis ; Identificació ; n ; Mé ; todos moleculares
• 刊名：Enfermedades Infecciosas y Microbiolog铆a Cl铆nica
• 出版年：2009
• 期刊代码：pca153_0213005X
• 类别：med
• 出版时间：November 2009
• 卷：27
• 期：9
• 页码：496-502
• 文件大小：307 K

Introduction

The m>Mycobacterium tuberculosism> complex includes the following species: m>Mycobacterium tuberculosis, Mycobacterium africanumm>, m>Mycobacterium bovis, Mycobacterium bovis-BCG, Mycobacterium microti, Mycobacterium capraem>, m>Mycobacterium pinnipedii,m> and m>Mycobacterium canettiim>. These species cause tuberculosis in humans and animals. Identification of mycobacterial strains has classically been performed by phenotype study. Over the last years, laboratories have developed several molecular techniques to differentiate between these species. The aim of this study is to evaluate these methods and develop a simple, fast, identification scheme.

Material and methods

We analyzed 251 strains randomly obtained from the strains studied in 2004, and 797 strains received by the Reference Laboratory between 2005 and 2007. Phenotype characterization of 4183 strains isolated during that period was done by studying the colony morphology, characteristics in culture, nitrate reduction, niacin accumulation, and growth in the presence of thiophen-2-carboxylic acid hydrazide 10 渭g/mL and pyrazinamide 50 渭g/mL. The molecular identification scheme designed was as follows: 1) m>gyrm>B PCR-RFLP with m>Rsam>I, m>Taqm>I or m>Sacm>II and m>hspm>65 RFLP/PCR with m>Hham>I., and 2) multiplex-PCR to determine the presence/absence of the RD9 and RD1 regions.

Results

The results showed 100%agreement between phenotype study and the molecular scheme.

Discussion

This molecular identification scheme is a simple and fast method, with 100%sensitivity and specificity, that can be implemented in most clinical laboratories at a low cost.