Thammasirirak Sompong, Siriteptawee Jaruwan, Sattayasai Nison, Indrakamhang Patchima, Araki Tomohiro
Department of Biochemistry, Faculty of Science, Khon Kaen University, Khon Kaen, Thailand.
Southeast Asian J Trop Med Public Health. 2003 Dec;34(4):751-7.
We established a new highly sensitive method, PCR-ELISA, for the dectection of Babesia bovis in cattle for farms in Thailand. The detection of around 2.4 x 10(-8)% parasitemia (equivalent to 1 infected erythrocyte per 2 ml) was achieved by PCR amplification followed by the ELISA detection of a biotin tagged gene. When comparing the sensitivity of PCR-ELISA with the microscopic method, our PCR-ELISA method is more sensitive than thin blood smears by at least 1,000 times. The established PCR-ELISA also showed high specificity to B. bovis with no cross reaction to other endemic parasites except for A. marginale. Regarding the detection threshold for B. bovis, the PCR-ELISA method could detect parasites inoculated into splenectomized calves at least 1 week earlier than the thin blood microscopic method. The PCR-ELISA method is a valuable screening technique for B. bovis and applicable for the routine detection of carrier states and automated analysis.
我们建立了一种新的高灵敏度方法——聚合酶链反应-酶联免疫吸附测定法（PCR-ELISA），用于检测泰国农场牛体内的牛巴贝斯虫。通过聚合酶链反应（PCR）扩增，随后采用酶联免疫吸附测定法检测生物素标记基因，实现了对约2.4×10⁻⁸%寄生虫血症（相当于每2毫升中有1个感染红细胞）的检测。当将PCR-ELISA的灵敏度与显微镜检查法进行比较时，我们的PCR-ELISA方法比薄血涂片法至少敏感1000倍。所建立的PCR-ELISA对牛巴贝斯虫也显示出高特异性，除边缘无浆体之外，与其他地方性寄生虫无交叉反应。关于牛巴贝斯虫的检测阈值，PCR-ELISA方法比薄血涂片显微镜检查法至少能提前1周检测到接种到脾切除小牛体内的寄生虫。PCR-ELISA方法是一种用于牛巴贝斯虫的有价值的筛查技术，适用于带虫状态的常规检测和自动化分析。
