High performance of an enzyme linked immunosorbent assay for American tegumentary leishmaniasis diagnosis with Leishmania (Viannia) braziliensis amastigotes membrane crude antigens
High performance of an enzyme linked immunosorbent assay for American tegumentary leishmaniasis diagnosis with Leishmania (Viannia) braziliensis amastigotes membrane crude antigens
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DOI：
10.1371/journal.pone.0232829
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发表时间：
2020-05-07
期刊：
影响因子：
3.7
通讯作者：
Marco, Jorge D.
中科院分区：
文献类型：
--
作者：
Bracamonte, Maria E.;Moya Alvarez, Agustin;Marco, Jorge D.
美国皮肤利什曼病（ATL）的诊断仍然需要设计更有效的工具。巴西利什曼原虫（Viannia）是阿根廷90%的ATL病例的病原体。结合目前的研究成果，设计了一种基于粗抗原的ELISA诊断方法.本研究纳入了99例诊断为ATL的受试者、27例非ATL受试者和84例来自非ATL流行区的供体。目前的ATL诊断基于四种技术，皮肤涂片显微镜检查（寄生虫学试验），PCR，利什曼宁皮肤试验和临床记录。我们获得了CA提取物的前鞭毛体和无鞭毛体巨噬细胞培养的地方性利什曼原虫物种循环在研究领域的不同酶组。从本地的主要酶模中提取的粗抗原。（五）与其他包括的物种相比，巴西利什曼原虫对抗利什曼原虫抗体表现出最高的反应性。无鞭毛体的CA比前鞭毛体高3.4倍。此外，无鞭毛体膜CA（MCA）的反应性比可溶性抗原高3.6倍。MCA-ELISA的敏感性和特异性分别为98%（CI = 94.7%~ 100%）和63.6%（CI = 53.9-73.1）。排除抗克氏锥虫反应血清后，特异性达98.4%（94.4 ~ 100），敏感性相似，阳性预测值（PV）为98.6%（94.6 ~ 100），阴性PV为96.3%（91.6 ~ 100）。MCA-ELISA结果的性能对最终诊断决策有很大贡献，因为无反应性血清学结果几乎可以排除疑似ATL，因为其高阴性PV。所开发的MCA-ELISA显示出高诊断性能，这使其成为ATL诊断、血清阳性率研究或监测治疗效果的良好候选者。
The diagnosis of American tegumentary leishmaniasis (ATL) still requires the design of more effective tools. Leishmania (Viannia) braziliensis is the causal agent of the 90% of Argentinean ATL cases. Considering the current knowledge, an ELISA based crude antigen (CA) for the diagnosis was designed. Ninety-nine subjects diagnosed as ATL, 27 as no-ATL, and 84 donors from non-ATL-endemic areas were included in this study. The current ATL diagnosis was based four techniques, dermal smear microscopic examination (parasitological test), PCR, Leishmanin skin test, and clinical records. We obtained CA extracts from promastigotes and amastigotes from macrophage cultures of different zymodemes of endemic Leishmania species circulating in the study area. Crude antigens from the 'local' main zymodeme of L. (V.) braziliensis showed the highest reactivity against anti-Leishmania antibodies compared to the other included species. The CA of amastigotes of this zymodeme was 3.4 fold more reactive than promastigotes one. Moreover, amastigote-membrane CA (MCA) were 3.6 fold more reactive than the soluble antigens. The MCA-ELISA reached a sensitivity and specificity of 98% (CI = 94.7%-100%) and 63.6% (53.9-73.1), respectively. When anti-Trypanosoma cruzi reactive sera were excluded, the specificity reached 98.4% (94.4-100), while the sensitivity was similar, with a positive predictive value (PV) of 98.6% (94.6-100) and negative PV of 96.3% (91.6-100). The performance of the MCA-ELISA results strongly contribute to the final diagnostic decision, since a non-reactive serological result almost discards the suspected ATL, because of its high negative PV. The developed MCA-ELISA showed a high diagnostic performance, which makes it a good candidate for ATL diagnosis, for seroprevalence studies, or for monitoring treatments efficacy.