| BACKGROUND AND OBJECTIVE:Tuberculosis,a disease of great antiquity holds lineage to saproptlytic soil organisms whose later introduction as a human pathogen likely coincided with the domestication of cattle approximately 10,000 years ago. In 1882 Robert Koch isoiated the causative agent, Mycobacterium tuberculosis from crushed lung tubercles. Since the tuberculosis has been found for one hundreds, the success of research, prevention, treatment with the disease makes people think that it will be exterminated by the end of the century.But now we find it is difficult to do.One-third of the world population is infected with Mycobacterium tuberculosis but only 5 to 10% of this population has a lifetime risk of developing active tuberculosis, either within 1 or 2 years after infection (primary tuberculosis) or thereafter (postprimary tuberculosis) When active tuberculosis develops, disease localization, severity, and outcome are highly variable. Tuberculosis may develop anywhere in the body, but usually presents as pulmonary infection, ranging from mild infiltration to chronic, cavitary, and severely destructive disease. The different manifestations of infection with M tuberculosis reflect the balance between the bacillus and host defense mechanisms, in which the quality of host defense determines outcome. Interleukin-1 (IL-1) , a cytokine with proinflammatory and fibrogenic effects., is known to be one of the pivotal mediators participating in aberrant immune responses in diffuse lung diseases. The most important members of the IL-1 family are the agonists IL-1α, IL-1β, and their naturally occurring inhibitor, IL-1 receptor antagonist (IL-1Ra). Genes encoding EL-1α, IL-1β, and IL-1Ra are clustered onchromosome 2q 13-21. Biallelic polymorphisms at positions IL-lP —511, and IL-lP -31 have been described, all representing a C/T single nucleotide polymorphism (SNP). The IL-IRa gene (1L-1RN) contains an 86-bp variable number tandem repeat (VNTR) polymorphism in intron 2. We have, therefore, performed a case-control study to investigate a plausible association between tuberculosis and the polymorphisms in the IL-lP, and IL-1 receptor antagonist (IL-IRa) genes.In the study we use two different methods to measure IL-1B gene polymorphisms to get the reliable result.SUBJECTS AND METHODS:l.Subjects:Peripheral Peripheral blood samples were obtained from 98 patients with tuberculosis (cases;male-female ratio, 1.09:1;range, 18-60 years)and 65 healthy controls (male-female ratio, 0.81:1;range, 18-60 years).2.IL-1B genotyping2.1 Technique of gene chip was used for the measurement of IL-1B gene polymorphism: Including DNA extraction, polymerase chain reaction(PCR),amplification ,hybridization,showing color.measurement and determining result.2.2 PCR-RFLP was used for the measurement of IL-1 B gene polymorphism:All genotyping of 98 cases and 65cdntrol samples wasperformed by DNA sequencing and polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). IL-1B polymorphisms were distinguished by PCR-RFLP, using the primer pairsand restriction enzymes. Amplification was performed in a volume of 25 u, 1, containing 2.5 y, 1 of 10 x PCR buffer (lOOmM Tris-HCL 15mMMgC12, and 500mMKCl, pH 8.3), 2.5mM each dNTP, 1 u, 1 each primer(10pmol/ \i 1),2.5U Taq DNA polymerase, and 3 |x 1 of genomic DNA. The thermocycling conditions were as follows: 94 |
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