Choi E J, Kang S W, Kweon C H, Jeong W S, Yoon Y D, Song H J
National Veterinary Research Institute, RDA, Anyang, Korea.
Korean J Parasitol. 1997 Jun;35(2):111-7. doi: 10.3347/kjp.1997.35.2.111.
Four separate pairs of oligonucleotide primers within the coding region in a T. sergenti 33-kDa surface protein gene were selected to detect T. sergenti by PCR. The specificity of PCR-amplified DNA was examined by digestion with restriction enzyme and Southern blot hybridization using T. sergenti p33 DNA probe. PCR appears to be specific for T. sergenti, without detectable signals from uninfected erythrocytes, uninfected bovine leukocytes and other hemoparasites, including A. marginale and B. ovata. Although 46 of 71 specimens (64.8%) from grazing cattle were microscopically positive, PCR in this study showed that 64 specimens (88.7%) were positive. Therefore, PCR proves a useful diagnostic tool for detecting T. sergenti-infected cattle. In addition, it is also revealed that PCR was significantly more sensitive than traditional microscopic examination using Giemsa's stain.
选择了布氏锥虫33-kDa表面蛋白基因编码区内的四对不同的寡核苷酸引物，通过聚合酶链反应（PCR）检测布氏锥虫。用限制性内切酶消化并使用布氏锥虫p33 DNA探针进行Southern印迹杂交，检查PCR扩增DNA的特异性。PCR似乎对布氏锥虫具有特异性，未感染的红细胞、未感染的牛白细胞以及其他血液寄生虫，包括边缘无浆体和卵形巴贝斯虫，均未检测到信号。尽管71份来自放牧牛的样本中有46份（64.8%）经显微镜检查呈阳性，但本研究中的PCR结果显示，有64份样本（88.7%）呈阳性。因此，PCR证明是检测布氏锥虫感染牛的一种有用诊断工具。此外，还发现PCR比使用吉姆萨染色的传统显微镜检查明显更敏感。
