Development of a genetic assay to distinguish between Leishmania viannia species on the basis of isoenzyme differences.
Development of a genetic assay to distinguish between Leishmania viannia species on the basis of isoenzyme differences.
复制标题
开发一种遗传测定法，根据同工酶差异区分维尼亚利什曼原虫物种。
DOI：
10.1086/500326
复制
发表时间：
2006
期刊：
影响因子：
--
通讯作者：
G. Matlashewski
中科院分区：
文献类型：
--
作者：
Wen;C. Miranda;J. Arévalo;M. Ndao;B. Ward;A. Llanos;G. Matlashewski
背景
拉丁美洲的表皮利什曼病主要由巴西利什曼原虫复合寄生虫引起。L.巴西利什曼原虫和秘鲁利什曼原虫是秘鲁的2种主要利什曼原虫。L.巴西利什曼病的毒性更强，因为它能引起皮肤粘膜利什曼病，称为埃斯彭迪亚，导致严重的面部破坏。早期鉴定引起初始皮肤感染的物种将大大有助于预防皮肤粘膜利什曼病，因为它将允许更积极的治疗和随访。然而，由于L. braziliensis和L.秘鲁，目前还没有简单的测定来区分这些物种。
方法
我们克隆了L. braziliensis和L.秘鲁。它是唯一已知的能够区分L. braziliensis和L.多位点酶电泳分析有趣的是，在L. braziliensis和L. Peruviana，导致氨基酸361处的氨基酸从苏氨酸变为精氨酸。本研究建立了一种特异性鉴别L. braziliensis或L.秘鲁。
结果
该检测方法与31个参考菌株进行了验证，这些参考菌株先前通过多位点酶电泳进行分型，成功应用于患者活检样本，并适用于实时聚合酶链反应检测。
结论
这种创新方法将新的遗传知识与多位点酶电泳的传统生化基础相结合，以更好地管理拉丁美洲的利什曼病。
BACKGROUND
Tegumentary leishmaniasis in Latin America is caused mainly by Leishmania viannia braziliensis complex parasites. L. braziliensis and Leishmania viannia peruviana are the 2 predominant Leishmania species in Peru. L. braziliensis is more virulent, because it can cause mucocutaneous leishmaniasis, known as espundia, that results in severe facial destruction. Early identification of the species that causes the initial cutaneous infection would greatly help to prevent mucocutaneous leishmaniasis, because it would allow more aggressive treatment and follow-up. However, because of the close genetic similarity of L. braziliensis and L. peruviana, there currently exists no simple assay to distinguish between these species.
METHODS
We cloned the mannose phosphate isomerase gene from both L. braziliensis and L. peruviana. It is the only known isoenzyme capable of differentiating between L. braziliensis and L. peruviana in multilocus enzyme electrophoresis. Interestingly, only a single nucleotide polymorphism was found between the mannose phosphate isomerase genes from L. braziliensis and L. peruviana, resulting in an amino acid change from threonine to arginine at amino acid 361. A polymerase chain reaction assay was developed to distinguish the single nucleotide polymorphism of the mannose phosphate isomerase gene to allow for the specific identification of L. braziliensis or L. peruviana.
RESULTS
This assay was validated with 31 reference strains that were previously typed by multilocus enzyme electrophoresis, successfully applied to patient biopsy samples, and adapted to a real-time polymerase chain reaction assay.
CONCLUSIONS
This innovative approach combines new genetic knowledge with traditional biochemical fundamentals of multilocus enzyme electrophoresis to better manage leishmaniasis in Latin America.