Microbiologie fondamentale et Pathogénicité, UMR 5234, Centre National de la Recherche Scientifique (CNRS), Université Bordeaux Segalen, Bordeaux, France.
PLoS One. 2013 Oct 25;8(10):e78565. doi: 10.1371/journal.pone.0078565. eCollection 2013.
Diagnosis of African animal trypanosomosis is vital to controlling this severe disease which hampers development across 10 million km(2) of Africa endemic to tsetse flies. Diagnosis at the point of treatment is currently dependent on parasite detection which is unreliable, and on clinical signs, which are common to several other prevalent bovine diseases.
METHODOLOGY/PRINCIPLE FINDINGS: the repeat sequence of the GM6 antigen of Trypanosoma vivax (TvGM6), a flagellar-associated protein, was analysed from several isolates of T. vivax and found to be almost identical despite the fact that T. vivax is known to have high genetic variation. The TvGM6 repeat was recombinantly expressed in E. coli and purified. An indirect ELISA for bovine sera based on this antigen was developed. The TvGM6 indirect ELISA had a sensitivity of 91.4% (95% CI: 91.3 to 91.6) in the period following 10 days post experimental infection with T. vivax, which decreased ten-fold to 9.1% (95% CI: 7.3 to 10.9) one month post treatment. With field sera from cattle infected with T. vivax from two locations in East and West Africa, 91.5% (95% CI: 83.2 to 99.5) sensitivity and 91.3% (95% CI: 78.9 to 93.1) specificity was obtained for the TvGM6 ELISA using the whole trypanosome lysate ELISA as a reference. For heterologous T. congolense field infections, the TvGM6 ELISA had a sensitivity of 85.1% (95% CI: 76.8 to 94.4).
CONCLUSION/SIGNIFICANCE: this study is the first to analyse the GM6 antigen of T. vivax and the first to test the GM6 antigen on a large collection of sera from experimentally and naturally infected cattle. This study demonstrates that the TvGM6 is an excellent candidate antigen for the development of a point-of-treatment test for diagnosis of T. vivax, and to a lesser extent T. congolense, African animal trypanosomosis in cattle.
诊断非洲动物锥虫病对于控制这种严重疾病至关重要,这种疾病在 1000 万平方公里的非洲地区流行,由采采蝇传播。目前,治疗点的诊断依赖于寄生虫检测,这种方法不可靠,也依赖于临床症状,但这些症状也常见于其他几种流行的牛病。
方法/原理发现:从几种锥虫 vivax(TvGM6)分离株中分析了 vivax 的 GM6 抗原的重复序列,尽管已知 vivax 具有高度遗传变异,但发现该重复序列几乎完全相同。TvGM6 重复序列在大肠杆菌中重组表达并纯化。在此抗原基础上开发了一种针对牛血清的间接 ELISA。在感染 vivax 后 10 天的实验感染后,TvGM6 间接 ELISA 的敏感性为 91.4%(95%置信区间:91.3%至 91.6%),一个月后治疗后敏感性降低十倍至 9.1%(95%置信区间:7.3%至 10.9%)。使用东非和西非两个地点感染 vivax 的牛的现场血清,使用整个锥虫裂解物 ELISA 作为参考,获得了 TvGM6 ELISA 的 91.5%(95%置信区间:83.2%至 99.5%)敏感性和 91.3%(95%置信区间:78.9%至 93.1%)特异性。对于异源 T. congolense 现场感染,TvGM6 ELISA 的敏感性为 85.1%(95%置信区间:76.8%至 94.4%)。
结论/意义:本研究首次分析了 vivax 的 GM6 抗原,也是首次在大量来自实验和自然感染牛的血清上检测 GM6 抗原。本研究表明,TvGM6 是一种优秀的候选抗原,可用于开发针对 vivax、在一定程度上也可用于 T. congolense 的治疗点诊断非洲动物锥虫病的检测。