Evaluation of a polymerase chain reaction assay for the diagnosis of bovine trypanosomiasis and epidemiological surveillance in Bolivia.
Evaluation of a polymerase chain reaction assay for the diagnosis of bovine trypanosomiasis and epidemiological surveillance in Bolivia.
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DOI：
10.1186/1475-9292-2-8
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发表时间：
2003-10-28
期刊：
影响因子：
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通讯作者：
Ribera Cuellar, Hugo
中科院分区：
文献类型：
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作者：
Gonzales, Jose Luis;Jones, Tudor W.;Ribera Cuellar, Hugo
背景：自1996年以来，玻利维亚就有牛锥虫病的零星暴发报道，当时T.间日疟原虫和T.首次通过寄生虫学手段鉴定出伊文思。然而，全面的流行病学资料，有关T。间日疟原虫和T.国内缺乏evansi。目前锥虫病的寄生虫学和血清学诊断方法在灵敏度或特异性方面存在重要局限性，这可能导致在流行病学研究中应用时数据不可靠。PCR检测是最近开发的一种方法，可能有助于克服寄生虫学和血清学检测的限制。因此，本研究的目的是评价PCR检测作为玻利维亚流行病学研究的诊断工具。结果如下：评估了用于诊断牛锥虫感染的PCR检测法检测从玻利维亚低地四个不同省份的牛采集的血斑样本中锥虫DNA的能力，并将结果与标准寄生虫学微量红细胞压积离心技术（MHCT）获得的结果进行了比较。染色涂片和血清学方法（T.卡凝集试验）。evansi（CATT）和T.间日疟原虫和T.刚果语）。Kappa一致性分析表明，PCR检测和寄生虫学方法的结果之间的显着协议，但没有协议时，PCR与血清学检测比较。从T.间日疟原虫涂片阳性动物经PCR检测为阴性，因此对PCR检测条件进行了修改，以尝试提高PCR和寄生虫学检测之间的一致性。模板DNA浓度的变化或使用替代引物组导致PCR检测率的提高，但并非所有寄生虫学阳性样品都能被PCR检测到。T.的PCR检测结果。间日疟原虫和T.将伊文思海吹综合分析结果与寄生虫学和血清学分析结果相结合，以提供关于采集样本的四个省的流行率的信息。结论：本研究确立了PCR作为流行病学研究诊断工具的有用性的证据，并证实牛锥虫病似乎是玻利维亚低地几个地区的地方病。
Background: Sporadic outbreaks of bovine trypanosomiasis have been reported in Bolivia since 1996 when T. vivax and T. evansi were identified for the first time by parasitological means. However, comprehensive epidemiological information concerning T. vivax and T. evansi in the country is lacking. Current parasitological and serological diagnostic methods for trypanosomiasis have important limitations either in their sensitivity or specificity, which can result in unreliable data when applied in epidemiological studies. PCR assays are a recently developed procedure that might help to overcome the constraints of parasitological and serological assays. Therefore, the objective of this study was to evaluate PCR assays as a diagnostic tool for epidemiological studies in Bolivia. Results: PCR assays for diagnosis of trypanosome infection in cattle were evaluated for their ability to detect trypanosome DNA in blood spots samples collected from cattle in four different provinces from the Bolivian lowlands and the results compared with those obtained with standard parasitological Micro Haematocrit Centrifugation Technique (MHCT) and stained smears and serological methods (Card Agglutination Test for T. evansi (CATT), and Antibody ELISAs for T. vivax and T. congolense). Kappa agreement analysis showed a significant agreement between PCR assays and results from parasitological methods but there was no agreement when PCR was compared with serological assays. Some samples from T. vivax smear positive animals were negative by PCR, therefore modifications to the PCR assay conditions were undertaken to try to improve agreement between PCR and parasitological assays. Changes in the template DNA concentration or the use of an alternative primer set resulted in improvements in the PCR detection rate, but not all the parasitologically positive samples were detected by PCR. Results from PCR assays for T. vivax and T. evansi were combined with results from parasitological and serological assays to provide information on prevalence rates for the four provinces from where the samples were obtained. Conclusion: The present study established evidence of the usefulness of PCR as diagnostic tool for epidemiological studies and confirmed that cattle trypanosomiasis appears to be endemic in several regions of the Bolivian lowlands.