Enzyme immunoassays (EIA) for serodiagnosis of human leptospirosis: specific IgG3/IgG1 isotyping may further inform diagnosis of acute disease.
Enzyme immunoassays (EIA) for serodiagnosis of human leptospirosis: specific IgG3/IgG1 isotyping may further inform diagnosis of acute disease.
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人钩端螺旋体病的酶免疫测定（EIA）：特定的IgG3/IgG1同种型可能会进一步为急性疾病的诊断提供信息。
DOI：
10.1371/journal.pntd.0010241
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发表时间：
2022-03
影响因子：
3.8
通讯作者：
Gomes-Solecki M
中科院分区：
文献类型：
--
作者：
Fortes-Gabriel E;Guedes MS;Shetty A;Gomes CK;Carreira T;Vieira ML;Esteves L;Mota-Vieira L;Gomes-Solecki M
费力的显微镜凝集试验（MAT）是钩端螺旋体病实验室诊断的金标准血清学试验。我们使用重组蛋白（rLigA、rLigB、rLipL 32）和来自8种钩端螺旋体血清型的全细胞提取物作为抗原开发了基于EIA的血清学检测，并评估了新检测方法在每个类别中对来自葡萄牙/PT（n = 143）和安哥拉/AO（n = 100）的MAT阳性（MAT+）人血清组的诊断性能。我们发现，重组蛋白rLigA、rLigB和rLipL 32的组合正确鉴定了来自安哥拉的罗安达省（LDA）和万博省（HBO）的临床和实验室确认的钩端螺旋体病（MAT+）患者血清中的抗原特异性IgG，灵敏度为92%，特异性约为97%（AUC 0.974）。将L. interrogans sv Copenhageni（LiC），L. kirschneri莫兹多克（LkM），L. borgpetersenii Arborea（LbA）和L. biflexa Patoc（LbP）准确识别了临床和实验室确诊的钩端螺旋体病（MAT+）患者，在安哥拉和葡萄牙的所有省份中，灵敏度为100%，特异性约为98%（AUC：AO/LDA/HBO为0.997，AO/HLA为1.000，PT/AZ为0.999，PT/LIS为1.000）。有趣的是，我们发现来自安哥拉的MAT+ IgG+血清具有显著更高的IgD存在，并且来自葡萄牙的MAT+ IgG+血清中IgG 3/IgG 1同种型显著增加。鉴于IgM/IgD类和IgG 3/IgG 1特异性同种型在感染的最早期产生，免疫球蛋白G同种型可用于诊断急性钩端螺旋体病。EIA检测的速度、易用性和准确性使其成为在致病性钩端螺旋体的循环血清型明确的地区筛查急性感染的费力和昂贵的MAT的极好替代品。钩端螺旋体病是一种潜在的致命性人畜共患疾病，由在受污染的水源和土壤中发现的细菌引起，在世界范围内分布。钩端螺旋体病的诊断是暂时基于在流行地区的医院就诊的患者的发热和肌痛的评估，并且由于缺乏负担得起的诊断测试，在世界大多数地区很少得到证实。我们利用酶免疫技术建立了一种高度敏感和特异的钩端螺旋体病血清学检测方法。在病原性钩端螺旋体的循环血清型明确的地区，多功能酶免疫测定法可以适应和操作的繁琐的金标准显微镜凝集试验的成本的一小部分，并应探索作为准确的钩端螺旋体病的血清学诊断工具。添加特异性免疫球蛋白G亚型（IgG 3/IgG 1）可进一步为急性钩端螺旋体病的诊断提供信息。
The laborious microscopic agglutination test (MAT) is the gold standard serologic test for laboratory diagnosis of leptospirosis. We developed EIA based serologic assays using recombinant proteins (rLigA, rLigB, rLipL32) and whole-cell extracts from eight Leptospira serovars as antigen and assessed the diagnostic performance of the new assay within each class, against MAT positive (MAT+) human sera panels from Portugal/PT (n = 143) and Angola/AO (n = 100). We found that a combination of recombinant proteins rLigA, rLigB and rLipL32 correctly identified antigen-specific IgG from patients with clinical and laboratory confirmed leptospirosis (MAT+) with 92% sensitivity and ~ 97% specificity (AUC 0.974) in serum from the provinces of Luanda (LDA) and Huambo (HBO) in Angola. A combination of whole cell extracts of L. interrogans sv Copenhageni (LiC), L. kirschneri Mozdok (LkM), L. borgpetersenii Arborea (LbA) and L. biflexa Patoc (LbP) accurately identified patients with clinical and laboratory confirmed leptospirosis (MAT+) with 100% sensitivity and ~ 98% specificity for all provinces of Angola and Portugal (AUC: 0.997 for AO/LDA/HBO, 1.000 for AO/HLA, 0.999 for PT/AZ and 1.000 for PT/LIS). Interestingly, we found that MAT+ IgG+ serum from Angola had a significantly higher presence of IgD and that IgG3/IgG1 isotypes were significantly increased in the MAT+ IgG+ serum from Portugal. Given that IgM/IgD class and IgG3/IgG1 specific isotypes are produced in the earliest course of infection, immunoglobulin G isotyping may be used to inform diagnosis of acute leptospirosis. The speed, ease of use and accuracy of EIA tests make them excellent alternatives to the laborious and expensive MAT for screening acute infection in areas where circulating serovars of pathogenic Leptospira are well defined. Leptospirosis is a potential fatal zoonotic disease with worldwide distribution caused by a bacterium found in contaminated sources of water and soil. Diagnosis of leptospirosis is tentatively based on evaluation of fever and myalgia in patients presenting at the hospital in areas of endemicity, and it is rarely confirmed in most parts of the world due to lack of affordable diagnostic tests. We developed a highly sensitive and specific serologic test for leptospirosis using ubiquitous enzyme immunoassay technology. In areas where circulating serovars of pathogenic Leptospira are well defined, versatile enzyme immunoassays can be adapted and operated at a fraction of the cost of the cumbersome gold standard Microscopic Agglutination Test and should be explored as accurate serodiagnostic tools for leptospirosis. Addition of specific immunoglobulin G isotyping (IgG3/IgG1) can further inform diagnosis of acute leptospirosis.
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从安哥拉捕获的啮齿动物的第一个分离株属于钩端螺旋体。
DOI：
10.4269/ajtmh.15-0027
发表时间：
2016-05-04
期刊：
The American journal of tropical medicine and hygiene
影响因子：
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2021-03-15
期刊：
The American journal of tropical medicine and hygiene
影响因子：
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Blacksell SD
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Kupferman, Tania;Coffee, Megan P.;Eckhardt, Benjamin J.
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