摘要
以边缘无浆体表面保护抗原的ｍｓｐｌβ基因设计和合成的１对２０ｍｅｒ寡核苷酸作为引物，用耐热的Ｔａｇ－ＤＮＡ聚合酶经５０个循环扩增边缘无浆体模板，扩增产物直接用凝胶电泳检测，并用标准分子量确定。结果表明，只有用边缘无浆体ＤＮＡ模板进行扩增时，扩增产物才能生成，而以血液原虫，如牛巴贝斯虫、双芽巴贝斯虫、锥虫和牛白细胞ＤＮＡ作模板扩增，无扩增产物生成。ＰＣＲ检测边缘无浆体的灵敏度可达约３００个无浆体的ＤＮＡ。
The Anaplasma marginale DNA was detected by using PCR technique. mspl β gene were used to design and synthesize a set of primers (each of 20 mer). The sequence selected for amplification consisted of 248 bp lying in the gene coding of mspl β. The standardized PCR condition were :Denaturation 94℃/40 s, annealing 45℃/1.5 min, extension 72℃/40 s, enzyme concentration 0.5 units. After 50 cycles of PCR by using the thermostable Tag DNA polymerase, the amplified products were directly detcted by gel electrophoresis and varified by molecular weight markers. The results showed that the amplified products were produced by amplification of the A.marginale DNA as template but not by amplification of other haemoparasites, such as Babesia bovis, Babesia bigemina, Trypanosoma and bovine WBC DNA (BWBC) as template under same conditions, indicating the specificity of the A.marginale PCR technique. The sensitivity of A.marginale PCR corresponds to the DNA of 300 A.marginale. The established A.marginale PCR technique provides a more sensitive and reliable methods for detection of carriers and study of this rickettsia.
出处
《中国兽医学报》
CAS
CSCD
北大核心
1997年第3期238-240,共3页
Chinese Journal of Veterinary Science
基金
国家劳动人事部流动调配司回国留学人员科研活动择优资助
关键词
边缘无浆体
牛
PCR
naplasma marginale
bovine
PCR
