CARD15基因StyⅠ酶切多态性与奶牛结核病易感性的相关性分析

Correlation Study of CARD15 Gene StyⅠPolymorphism and Susceptibility of Tuberculosis in Dairy Cows

本试验旨在探讨CARD15基因StyⅠ酶切位点多态性与云南高原奶牛结核病易感性的关联。选择云南昆明、玉溪、大理等地结核病阳性奶牛103头,结核病阴性奶牛98头,共201头为研究对象,应用创造酶切位点原理设计引物,通过PCR-RFLP与PCR直接测序法相结合的检测方法,获得CARD15基因StyⅠ酶切位点的基因型及等位基因频率,分析该位点与云南高原奶牛结核病易感性的关系。结果显示,CARD15基因的StyⅠ酶切位点存在AA、AG、GG 3种基因型,在结核病例组中AA、AG、GG基因型频率为0.03、0.42和0.55,A、G等位基因频率为0.24和0.76;在结核对照组中AA、AG、GG基因型频率为0.01、0.05和0.94,A、G等位基因频率为0.04和0.96;对多态性位点的遗传模式进行分析,最佳遗传模式为显性遗传(dominant),在此遗传模式下基因型AG-AA在病例组的频率44.7%,明显高于对照组的频率6.1%,OR(95%CI)=0.08(OR<1),在病例组与对照组的分布之间有极显著差异(P<0.0001)。结果表明,云南高原奶牛CARD15基因StyⅠ酶切位点多态性与结核病的发生有显著关联性,且基因型AG-AA可能是结核易感型。

The aim of this study was to investigate the association between CARD15 gene Sty I polymorphism and susceptibility to tuberculosis （TB） of dairy cows from Yunnan plateau. A total of 201 dairy cows （103 cases and 98 controls） from Kunming,Yuxi and Dali were used in this study. We designed a specific primer pair for PCR-RFLP analysis. By using PCR-RFLP genoty- ping and PCR sequencing, We analysized allele frequency and correlation between polymorphism of CARD15 gene and susceptibility to bovine tuberculosis in Yunnan province. The results showed that there were three genotypes in Sty I enzyme site, AA, AG and GG. In cases, the genotypefrequencies of AA, AG and GG were 0.03,0.42 and 0. 55, respectively, while in controls were 0.01,0.05 and 0.94,and the allele frequencies of A and G were 0.24 and 0.76 in cases,while were 0. 04 and 0. 96 in controls. Then analysizing the genetic pattern of polymorphic loci, we found that the best genetic model was dominant and the frequency of AG-AA in cases （44.7%） was apparently higher than that in controls （6.1%） （OR（95% CI）=0.08 （OR〈1））,so extreme- ly significant difference existed between cases and controls （P%0. 0001）. The polymorphism of Sty I enzyme cut site in CARD15 gene was significantly associated with the susceptibility to tubercu- losis of Yunnan plateau dairy cows, and AG-AA might be the susceptible genotype to tuberculosis.