Fernández D, González-Baradat B, Eleizalde M, González-Marcano E, Perrone T, Mendoza M
Universidad Nacional Experimental Simón Rodríguez, Instituto de Estudios Científicos y Tecnológicos, Centro de Estudios Biomédicos y Veterinarios, Apartado Postal 47925, Caracas 1041A, Venezuela.
Exp Parasitol. 2009 Jan;121(1):1-7. doi: 10.1016/j.exppara.2008.09.013. Epub 2008 Sep 26.
Trypanosoma evansi is the causative agent of equine trypanosomosis, disease that affects horse's productivity and health. Parasitological and molecular methods are mostly used to detect the infection. The aim of this work was evaluate PCR sensitivity to detect T. evansi using the primers 21/22-mer, ITS1, ESAG 6/7 and TBR 1/2 designed from repetitive (multicopies) genomic sequences. The results were compare with two parasitological tests in mice, micro-haematocrite centrifugation technique and direct microscopic examination. The results shows (a) that the minimum amount of DNA from blood of highly parasitaemic mice that was detectable by PCR was 0.001 ng, using the ESAG6/7 and TBR1/2 primer, (b) using TBR1/2 primer for parasites purified could detect 0.000001 ng and (c) in the prepatent period PCR detect the presence of parasites earlier than parasitological techniques. Nevertheless, the percentage of detection for PCR varies depending on primer employed with 60% and 66% for ITS1 and 21/22-mer, and 80% for ESAG6/7 and TBR1/2. Consequently, TBR1/2 and ESAG6/7 were the best primers to monitor T. evansi infections in mice. For epidemiological application, such comparative evaluation should be made for detection of T. evansi in livestock such as horses.
伊氏锥虫是马锥虫病的病原体,该疾病会影响马的生产力和健康。寄生虫学和分子方法大多用于检测感染情况。这项工作的目的是评估聚合酶链反应(PCR)检测伊氏锥虫的敏感性,使用从重复(多拷贝)基因组序列设计的21/22-mer、ITS1、ESAG 6/7和TBR 1/2引物。将结果与小鼠的两种寄生虫学检测方法进行比较,即微量血细胞比容离心技术和直接显微镜检查。结果显示:(a)使用ESAG6/7和TBR1/2引物,PCR可检测到的高度寄生虫血症小鼠血液中DNA的最小量为0.001纳克;(b)使用TBR1/2引物对纯化的寄生虫可检测到0.000001纳克;(c)在潜伏期,PCR比寄生虫学技术更早检测到寄生虫的存在。然而,PCR的检测百分比因所用引物而异,ITS1和21/22-mer为60%和66%,ESAG6/7和TBR1/2为80%。因此,TBR1/2和ESAG6/7是监测小鼠伊氏锥虫感染的最佳引物。对于流行病学应用,应针对马等家畜中伊氏锥虫的检测进行此类比较评估。