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用于检测马提尼克利什曼原虫感染的抗体的驱动蛋白39、热休克蛋白70、热休克蛋白83和糖蛋白63重组蛋白的比较

Comparison of Recombinant Proteins of Kinesin 39, Heat Shock Protein 70, Heat Shock Protein 83, and Glycoprotein 63 for Antibody Detection of Leishmania martiniquensis Infection.

作者信息

Siripattanapipong Suradej, Kato Hirotomo, Tan-Ariya Peerapan, Mungthin Mathirut, Leelayoova Saovanee

机构信息

Department of Microbiology, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand.

Division of Medical Zoology, Department of Infection and Immunity, Jichi Medical University, Tochigi, 329-0498, Japan.

出版信息

J Eukaryot Microbiol. 2017 Nov;64(6):820-828. doi: 10.1111/jeu.12415. Epub 2017 Apr 27.

Abstract

Leishmania martiniquensis, a zoonotic hemoflagellate, is a causative agent of cutaneous (CL) and visceral leishmaniasis (VL) among humans and animals. This organism, first reported in Martinique Island, now has become an emerging infectious agent in Thailand. Symptomatic cases of L. martiniquensis infection among humans have continuously increased. In the meantime, asymptomatic infection of this novel species has seriously created national public health awareness and concern to prevent and control disease transmission. The unsuccessful serological test using the commercial rK39 dipstick based on antigen from Leishmania donovani to detect the antibodies against VL among infected Thai patients has encouraged us to further explore a new sensitive and specific antigenic epitope. In this study, we determined the sequences and expressed recombinant proteins of kinesin 39 (k39), heat shock protein 70 (hsp70), heat shock protein 83 (hsp83), and glycoprotein 63 (gp63) of L. martiniquensis to evaluate the diagnostic efficiency to detect antibodies against L. martiniquensis in patient sera. The preliminary results from western blot analysis have suggested that K39 is the most sensitive recombinant protein to detect L. martiniquensis. Moreover, this recombinant protein reacts with antibodies against L. donovani and Leishmania infantum, making it a promising antigen for further development of a universal rapid diagnostic tool for VL.

摘要

马提尼克利什曼原虫是一种人畜共患的血鞭毛虫,是人类和动物皮肤利什曼病(CL)和内脏利什曼病(VL)的病原体。这种生物体最早在马提尼克岛被报道,现在已成为泰国一种新出现的感染源。人类中马提尼克利什曼原虫感染的症状性病例持续增加。与此同时,这种新物种的无症状感染严重引起了国家公众对预防和控制疾病传播的健康意识和关注。基于来自杜氏利什曼原虫的抗原使用商业rK39试纸条进行的血清学检测未能成功检测感染泰国患者中针对VL的抗体,这促使我们进一步探索一种新的敏感且特异的抗原表位。在本研究中,我们测定了马提尼克利什曼原虫的驱动蛋白39(k39)、热休克蛋白70(hsp70)、热休克蛋白83(hsp83)和糖蛋白63(gp63)的序列并表达了重组蛋白,以评估检测患者血清中针对马提尼克利什曼原虫抗体的诊断效率。蛋白质印迹分析的初步结果表明,K39是检测马提尼克利什曼原虫最敏感的重组蛋白。此外,这种重组蛋白与针对杜氏利什曼原虫和婴儿利什曼原虫的抗体发生反应,使其成为进一步开发VL通用快速诊断工具的有前景的抗原。

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