| Objective: To investigate the distribution of transporter of antigen peptides(TAP)genes TAP1333and TAP1637 in normal human being and allergic rhinitis(AR) patients,and to explore the association of these gene polymorphisms with allergic rhinitis.Methods:①Detection of allergen-specific IgE. Peripheral venous blood (about 2 ml) was drawn from each individual and centrifugate those blood, and segregate the serum. In this study the AD in vitro diagnosis box was used to detect the serum levels of allergen-specific IgE antibodies(9 items). The level of strength about positive results were determined on the basis of the coloration shade of test paper,and separately to express by +,++, +++.②DNA extraction. Heparinized peripheral venous blood (about 2 mL) was drawn from each individual. Genomic DNA was extracted from leukocytes in the collected blood with the TIANamp blood DNA kit. Using UV spectrophotometer to determinate the purity and concentration of DNA samples,and record the OD value of 260nm,280nm wavelength.③Primer synthesis. Four primers were used for each dimorphic site of TAP1333 and TAP1 637. Including a pair of common primers and two specific primers. Primers were designed as described by Powis et al.[4]④PCR Amplification. According to the different allelotypes, We have designed three different primers combinations and add to A,B,C three tubes. The amplification products of each tube on behalf of the same one subject at the different primer extension for the reaction system.⑤Agarose Gel Electrophoresis. The mixture of reaction products were separated on a 1.5% agarose gel stained with ethidium bromide. To observe and analysis the results by the ultraviolet lamp.⑥Statistical analysis. All statistical analyses were performed with SPSS 11.0. A chi-square test was used to compare the distribution of TAP genotypes between control and AR groups. Differences were significant if p<0.05.Results: 1 Detection of sIgE: In the sequence of all the inhalant allergens, the highest positive rates was dermatophagoides pteronyssinus,the next were gramineae polenand and epidermis of animal.2 The purity and concentration of DNA samples: The ratio of OD260/OD280 were between 1.5 ~ 2.0. Some samples may exist RNA or protein contamination, but does not affect the results of PCR amplification.3 Result of PCR. Genotyping of the TAP1333 was done as follows: Ile/Val if 241, 351 and 533 bp bands were visible, Ile/Ile if 241 and 533 bp bands were visible, and Val/Val if 351 and 533 bp bands were visible . Genotyping of the TAP1637 was done as follows: Asp/Gly if 180, 307 and 429 bp bands were visible,Asp/Asp if 307 and 429 bp bands were visible, andGly/Gly if 180 and 429 bp bands were visible.Three TAP1 genotypes (Ile/Ile, Ile/Val and Val/Val) were found at codon 333. Two TAP1 genotypes (Asp/Asp, Asp/Gly) were seen at codon 637.The Gly /Gly genotype was not found in both AR and controls.4 Result of statistical analysis: II and DD were the most common genotype of the TAPl codon 333 and 637 in both the AR group and control group. The TAP1333 was found have statistical significance as compared with the control group(P<0.05). no statistical significance was obtained in the latter(P> 0.05). I and D were the most common allele of the TAPl codon 333 and 637 in both the AR group and control group. The TAP1333 was found have statistical significance as compared with the control group(P<0.05). no statistical significance was obtained in the latter(P> 0.05).Conclusions:In conclusion,this study showed a meaningful association between AR and TAP1333 polymorphism, but not TAP1637. Therefore, it is suggested that a TAP1333 gene polymorphism may be an important factor in AR pathogenesis. TAP as susceptibility/protective gene of AR affected by the objective factors such as selection of patients,different population, regional environment.And its specific genetic significance has to be further clarified.
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