IDENTIFICATION OF BIOMARKERS FOR TUBERCULOSIS IN PATIENTS WITH TYPE 2 DIABETES MELLITUS
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1
IMSS, UNIDAD DE INVESTIGACION BIOMEDICA DE ZACATECAS, Mexico
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2
SERVICIOS DE SALUD DE ZACATECAS, COORDINACION DE INVESTIGACION, Mexico
Tuberculosis by Mycobacterium tuberculosis infection is a major cause of morbidity and mortality worldwide. In humans, comorbidities with HIV (human immunodeficiency virus) and Type 2 diabetes mellitus (DM2) increase the susceptibility of tuberculosis (TB). In Mexico, there is an increased risk to develop TB in DM2 patients (7 times more compared to not DM2 patients). The comorbidity of TB-DM2 in developing countries is increasing the magnitude of this problem. According to this, it is an urgent necessity to identify biomarkers that are capable to discriminate between patients DM2/TB from latent tuberculosis infected (LTB) with DM2 who are in higher risk in developing tuberculosis. For this purpose, a microarray analysis was performed to identify a transcriptional signature that distinguishes DM2 active tuberculosis individuals (TBA/DM2) from DM2 latent infected (LTB/DM2) and from DM2 non-TB infected (CTR/DM2). Using a custom Combimatrix Microarray platform (IMSS4X2K); total mRNA from whole peripheral blood was obtained by PaxGene RNA isolation kit (Qiagen Inc., USA) from the 3 described groups; labelled cRNA was hybridized to the microarray chip. Cluster analysis and statistical microarray analysis was done using Cluster, Treeview and SAM software to identify differentially expressed genes. An unsupervised analysis of the microarray data identified the 2 fold up-regulated genes with a q-value<1, in the TB/DM2 group compared to LTB/DM2 and CTR/DM2 groups. RT-qPCR methodology was used to validate a gene subset from microarray results. The comparisons between groups were analysed using the Graph Pad Prism software 5.0 and Graph Pad InStat (San Diego, Ca., USA), and LDA and ROC curves were performed using the Statistical Package for Social Sciences v.16.0 (Chicago, SPSS Inc.).
A transcriptome of 13 up-regulated genes (more than five fold increase) was identified. Ten of these genes were validated by qPCR amplification in the 3 study groups. Four genes showed significant differences between TB/DM2 group compared to LTB/DM2 and CTR/DM2 groups. These 4 genes conserve their differences when we merge the CTR/DM2 and the LTB/DM2 as one (NoTB/DM2) and compare it with the TB/DM2 group. To identify the best biomarkers with the highest discriminatory power, we performed a linear discriminant analysis (LDA) and ROC curves. ROC curves were used to identify the cut-off value for the best sensitivity and specificity values. ROC curves demonstrate that biomarkers have sensitivities between 80 to 90% and specificities between 85 to 95%. To increase the diagnostic capacity of these BM´s, we performed a serial and parallel multiple tests. The highest sensitivity and specificity for TB diagnosis in patients with DM2 was found using only three BM. In a serial test, the combination of BM1 and BM2 gave 100% sensitivity and 85% specificity. When BM1 and BM3 were combined, in a parallel test a 90% sensitivity and 100% specificity was achieved
A specific transcriptome associated to Tuberculosis in subjects with DM2 was identified. A subset of 3 biomarkers is able to discriminate in a serial and parallel tests patients with TB/DM2 from subjects with LTB/DM2 or CTR/DM2.
Acknowledgements
THIS WORK WAS SUPPORTED BY CONACYT, MEXICO GRANT 215380, PDCPN2013-01. JAEM IS A SCHOLAR OF FUNDACION IMSS
Keywords:
Tuberculosis,
biomarkers,
Diagnosis, Differential,
Infection,
Microarrays
Conference:
IMMUNOCOLOMBIA2015 - 11th Congress of the Latin American Association of Immunology - 10o. Congreso de la Asociación Colombiana de Alergia, Asma e Inmunología, Medellin, Colombia, 13 Oct - 16 Oct, 2015.
Presentation Type:
Poster Presentation
Topic:
Infectious and parasitic diseases
Citation:
ENCISO-MORENO
J,
MACIAS-SEGURA
N,
SERRANO- ESCOBEDO
C and
GANDARA-JASSO
B
(2015). IDENTIFICATION OF BIOMARKERS FOR TUBERCULOSIS IN PATIENTS WITH TYPE 2 DIABETES MELLITUS.
Front. Immunol.
Conference Abstract:
IMMUNOCOLOMBIA2015 - 11th Congress of the Latin American Association of Immunology - 10o. Congreso de la Asociación Colombiana de Alergia, Asma e Inmunología.
doi: 10.3389/conf.fimmu.2015.05.00200
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Received:
15 May 2015;
Published Online:
14 Sep 2015.
*
Correspondence:
PhD. JOSE ANTONIO ENCISO-MORENO, IMSS, UNIDAD DE INVESTIGACION BIOMEDICA DE ZACATECAS, ZACATECAS, ZACATECAS, 98000, Mexico, enciso_2000@yahoo.com