Cytokines Interleukin 4 (IL-4) and Interleukin 10 (IL-10) Gene Polymorphisms as Potential Host Susceptibility Factors in Virus-Induced Encephalitis

Yu, Ying; Chen, Ying; Wang, Feng-Ling; Sun, Jing; Li, Hai-Jun; Liu, Jia-Ming

The PCR primers were designed by Primer Premier 5.0 and synthesized by Shanghai Sangon BioTech Co., Ltd. (Shanghai, China). The forward and reverse primers of IL-4 and IL-10 gene polymorphisms are shown in Table 1 and IL-10 rs1800871/rsl800872 shared a pair of primers. The total volume of PCR reaction system was 50 μl, including 0.5 μl of DNA template, 5 μl of forward primer and reverse primer, respectively, 25 μl of PCR-pfu mix enzyme, and 14.5 μl of water. After DNA was dissolved, 5-μl samples (concentration >0.1 μg/μl) were assessed using an ultraviolet spectrophotometer on the basis of the ratio (260/280) of the range between 1.8 to 2.1. The PCR reaction conditions were as follows: pre-denaturation at 94°C for 45 s, annealing at 60°C for 45 s, extension at 72°C for 45 s, 39 cycles, and extension at 72°C for 5 min by proper adjustment of annealing temperature according to primer synthesis report.