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目的:了解我国北方部分地区宿主动物感染土拉弗菌状况和基因型别。方法:鼠夹诱捕鼠,从鼠脾组织提取DNA,应用巢式PCR检测土拉弗菌,计算并比较各地区及各鼠种阳性率。阳性标本用型特异性引物及短串联重复序列(SSTR9、SSTR16)引物扩增两区域并测序,用C lustalX(5.0)和DNA C lub软件对序列进行比较分析。结果:共捕获鼠421只,分属14种鼠种,其中7个鼠种20份标本PCR检测阳性,总阳性率为4.75%。吉林、新疆、黑龙江、内蒙古和浙江的阳性率分别为11.65%,10.00%,6.54%,1.76%和0%。地区间阳性率差异显著(χ2=20.90,P=0.0003);不同鼠种间阳性率未见差异(χ2=11.82,P=0.066)。对20份扩增阳性标本进行型特异性扩增及SSTR9、SSTR16序列测定并对比分析,结果显示均为土拉弗菌B亚型,但型内差别较大,标本及菌株相互比较均显示基因多态性程度高且地区间有交叉。结论:我国北方部分地区可能依然存在B亚型土拉弗菌型感染(宿主动物)情况,且其基因型内差异较大。
OBJECTIVE: To understand the status and genotype of Tetrastigma infection in host animals in some parts of northern China. Methods: Rattrap traps were used to extract DNA from spleen tissue of rats. Nephroda was used to detect Tetrara toxins. The positive rate of each species was calculated and compared. The positive samples were amplified using two types of primers and specific primers and short tandem repeat (SSTR9, SSTR16) primers and sequenced. The sequences were analyzed by C lustalX (5.0) and DNA C lub software. Results: A total of 421 mice were captured, belonging to 14 kinds of mice, of which 7 specimens of 20 samples PCR positive, the total positive rate was 4.75%. The positive rates of Jilin, Xinjiang, Heilongjiang, Inner Mongolia and Zhejiang were 11.65%, 10.00%, 6.54%, 1.76% and 0% respectively. There was significant difference between the two groups (χ2 = 20.90, P = 0.0003). There was no difference in the positive rate among different species (χ2 = 11.82, P = 0.066). Type-specific amplification and SSTR9 and SSTR16 sequencing of 20 positive amplification samples were performed and analyzed. The results showed that all of them were T subtype B. However, High degree of polymorphism and cross between regions. Conclusion: There may still be cases of Tetragenotype B infection (host animals) in some parts of northern China, and the genotypes of these strains are quite different.