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用于犬钩端螺旋体感染诊断的间接酶联免疫吸附测定(ELISA)开发中的富含亮氨酸重复序列蛋白及衍生肽

Leucine-Rich Repeat Proteins and Derived Peptides in an Indirect ELISA Development for the Diagnosis of Canine Leptospiral Infections.

作者信息

Sripattanakul Sineenat, Prapong Teerasak, Kamlangdee Attapon, Katzenmeier Gerd, Haltrich Dietmar, Hongprayoon Ratchanee, Prapong Siriwan

机构信息

The Interdisciplinary Graduate Program in Genetic Engineering, The Graduate School, Kasetsart University, Bangkok 10900, Thailand.

Department of Physiology, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand.

出版信息

Trop Med Infect Dis. 2022 Oct 17;7(10):311. doi: 10.3390/tropicalmed7100311.

Abstract

Domestic and stray dogs can be frequently infected by , and thus may represent a source for transmission of this zoonotic disease in Thailand. Here, we have used peptides derived from a recombinant leucine-rich repeat (LRR) protein of , rKU_Sej_LRR_2012M, for the development of an indirect enzyme-linked immunosorbent assay (ELISA) aimed at detecting antibodies against , , and , the three major seroprevalences in Thai dogs. The rKU_Sej_LRR_2012M protein is recognized by hyperimmune sera against several leptospiral serovars. The epitope peptides of the rKU_Sej_LRR_2012M showed binding affinities with lower IC values than peptides of known antigenic protein LipL32. Four peptides, 2012-3T, 2012-4B, 2012-5B and pool 2012-B, were specifically recognized by rabbit hyperimmune sera against nine serovars from three spp. The indirect peptide-based ELISAs with these four peptides were evaluated with the LipL32 ELISA by using a receiver-operator curve (ROC) analysis. All peptides had an area under the curve of ROC (AUC) greater than 0.8, and the sum of sensitivity and specificity for each peptide was greater than 1.5. The degree of agreement of 2012-3T and pool 2012-B and 2012-4B and 2012-5B peptides were in moderate-to-good levels with kappa values of 0.41-0.60 and 0.61-0.80, when compared with LipL32, respectively. This finding would suggest an excellent capability of the 2012-4B and 2012-5B peptide-based ELISAs assay for the diagnosis of canine leptospiral infections.

摘要

家犬和流浪狗经常会感染[病原体名称未给出],因此可能是泰国这种人畜共患病传播的一个源头。在此,我们使用了源自重组富含亮氨酸重复序列(LRR)蛋白rKU_Sej_LRR_2012M的肽段,来开发一种间接酶联免疫吸附测定(ELISA),旨在检测泰国犬类针对[病原体名称未给出]、[病原体名称未给出]和[病原体名称未给出]这三种主要血清流行株的抗体。rKU_Sej_LRR_2012M蛋白可被针对几种钩端螺旋体血清型的超免疫血清识别。rKU_Sej_LRR_2012M的表位肽段与已知抗原蛋白LipL32的肽段相比,显示出结合亲和力且IC值更低。四种肽段,即2012 - 3T、2012 - 4B、2012 - 5B和混合肽段2012 - B,可被针对来自三个[钩端螺旋体属物种名称未给出]物种的九种血清型的兔超免疫血清特异性识别。通过使用受试者工作特征曲线(ROC)分析,对使用这四种肽段的基于肽段的间接ELISA与LipL32 ELISA进行了评估。所有肽段的ROC曲线下面积(AUC)均大于0.8,并且每种肽段的敏感性和特异性之和大于1.5。与LipL32相比,2012 - 3T和混合肽段2012 - B以及2012 - 4B和2012 - 5B肽段的一致性程度处于中等至良好水平,kappa值分别为0.41 - 0.60和0.61 - 0.80。这一发现表明基于2012 - 4B和2012 - 5B肽段的ELISA检测法在诊断犬类钩端螺旋体感染方面具有出色的能力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9e23/9610812/c5b1f4967657/tropicalmed-07-00311-g001.jpg

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