H. J. Chang, M. Y. Huang, C. S. Yeh, C. C. Chen, M. J. Yang, C. S

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Rapid diagnosis of tuberculosis directly from clinical specimens using a gene chip H.J. Chang, M.Y. Huang, C.S. Yeh, C.C. Chen, M.J. Yang, C.S. Sun, C.K. Lee, S.R. Lin Clinical Microbiology and Infection Volume 16, Issue 8, Pages 1090-1096 (August 2010) DOI: 10.1111/j.1469-0691.2009.03045.x Copyright © 2010 European Society of Clinical Infectious Diseases Terms and Conditions

FIG. 1 PCR products from Mycobacterium samples obtained with with hsp65 and IS6110 probes and results of BstE-generated and HaeIII-generated restriction fragment length polymorphism (RFLP) patterns. Two sets of oligonucleotide primers were used to amplify target DNA from the multiple-copy hsp65 gene and IS6110. A 440-bp fragment of hsp65 was amplified by PCR using primers TB11 and TB12. A 542-bp fragment of IS6110 was amplified by PCR using primers MT1007 and MT1008. The PCR products were analysed by electrophoresis on a 3% agarose gel stained with ethidium bromide, and examined for the presence of 440-bp and 542-bp fragments under UV irradiation. (a) Cases A, B, C, D and E were hsp65-positive and IS6110-positive; cases F and G were hsp65-positive and IS6110-negative; case H was hsp65-negative and IS6110-negative. (b) PCR products were digested with the restriction enzymes BstE and HaeIII in order to identify Mycobacterium tuberculosis complex (MTBC) and non-tuberculous mycobacteria (NTM) on the basis of the RFLP patterns. Cases A, B, C, D and E were confirmed MTBC. Cases F and G were NTM. Case H involved another species. Clinical Microbiology and Infection 2010 16, 1090-1096DOI: (10.1111/j.1469-0691.2009.03045.x) Copyright © 2010 European Society of Clinical Infectious Diseases Terms and Conditions

FIG. 2 Array table of the gene chip and gene expression pattern in a Mycobacterium tuberculosis sample. (a) Schematic representation of a membrane array with 13 M. tuberculosis-specific target genes, one housekeeping gene (β-actin), and a blank control. The positions of 13 M. tuberculosis-specific target genes (i.e. hsp65, Rv0577, TbD1, Rv3120, Rv2073c, Rv1970, Rv3875, Rv0186, Rv0124, Rv3347, Rv1510, mtp40 and mpb83), as well as the correlation between the blank and positive control (β-actin) in the nylon membrane, are indicated by spots. The relevant positions are red (β-actin), yellow (blank), and blue, green and purple (M. tuberculosis-specific target genes). (b) Expression patterns of M. tuberculosis-specific target genes in one sample of M. tuberculosis; the array results were as follows: hsp65, +; Rv05 77, +; TbD1, –; Rv3120, +; Rv2073c, +; Rv1970, +; Rv3875, +; Rv0186, +; Rv0124, +; Rv3347, +; Rv1510, +; mtp40, +; mpb83, +; β-actin, +. Clinical Microbiology and Infection 2010 16, 1090-1096DOI: (10.1111/j.1469-0691.2009.03045.x) Copyright © 2010 European Society of Clinical Infectious Diseases Terms and Conditions