A novel single-tier serologic test to diagnose all stages of Lyme disease.

IF 5.4 2区 医学 Q1 MICROBIOLOGY
Journal of Clinical Microbiology Pub Date : 2025-09-10 Epub Date: 2025-08-20 DOI:10.1128/jcm.00483-25
Andrew E Levin, Gary P Wormser, Elizabeth J Horn, Nadezhda Karaseva, Drew Miller, Hunter Kellogg
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引用次数: 0

Abstract

Lyme disease, a bacterial zoonosis, is the most commonly reported vector-borne disease in the United States. Laboratory diagnosis has relied on a two-tier serologic approach, originally comprising an ELISA, or another first-tier assay, followed by separate IgG and IgM immunoblots to confirm a positive first-tier result. This standard two-tier testing (STTT) approach provides high specificity, but at the cost of low sensitivity in early Lyme disease. Recent studies have shown that a modified two-tier (MTTT) testing approach, in which a second ELISA replaces the immunoblot, can provide an increase in test sensitivity without a loss of specificity. Nevertheless, neither STTT nor MTTT is considered sensitive enough for diagnosing patients with erythema migrans, the most common clinical manifestation of early Lyme disease. We have developed a novel ELISA methodology termed "Hybrid Lyme ELISA" for single-tier Lyme antibody detection, which relies on the simultaneous binding of individual antibody molecules to the Borrelia burgdorferi surface protein VlsE and to the C6 peptide derived from it. This dual binding requirement builds exceptionally high specificity into the assay, eliminating the majority of non-specific antibody interactions. In this study, the single-tier Hybrid Lyme ELISA was shown to provide greater sensitivity, but with equivalent specificity, to both STTT and MTTT. In addition, given the >90% sensitivity of the Hybrid Lyme ELISA in patients with erythema migrans, this assay may not only transform serologic testing from two-step to single-step testing, but may also provide a means for the first time to diagnose patients with erythema migrans.IMPORTANCEThe diagnosis of Lyme disease, a tick-borne spirochetal infection caused by Borrelia burgdorferi sensu lato, is subject to two major limitations: the need for a two-tier serologic testing algorithm to provide adequate specificity, and the low sensitivity of this algorithm in practice for detection of early Lyme disease manifesting with the erythema migrans skin lesion, the most common clinical manifestation. This study presents the first description of a new assay, the Hybrid Lyme ELISA, which demonstrates sensitivity high enough to potentially diagnose over 90% of patients with erythema migrans, and specificity high enough to preclude the need for a second-tier test. These test characteristics suggest the potential for the Hybrid Lyme ELISA to be the first single-tier serologic test suitable for laboratory diagnosis of all stages of Lyme disease.

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一种新的单层血清学测试来诊断莱姆病的所有阶段。
莱姆病是一种细菌性人畜共患病,是美国最常见的媒介传播疾病。实验室诊断依赖于两层血清学方法,最初包括ELISA或另一层测定,然后是单独的IgG和IgM免疫印迹,以确认第一层阳性结果。这种标准的两层检测(STTT)方法提供了高特异性,但代价是早期莱姆病的低敏感性。最近的研究表明,一种改进的两层(MTTT)检测方法,即用第二次ELISA代替免疫印迹,可以在不丧失特异性的情况下提高检测灵敏度。然而,对于早期莱姆病最常见的临床表现——移行性红斑的诊断,STTT和MTTT都不够敏感。我们开发了一种新的ELISA方法,称为“混合莱姆病ELISA”,用于单层莱姆病抗体检测,它依赖于单个抗体分子与伯氏疏螺旋体表面蛋白VlsE及其衍生的C6肽的同时结合。这种双重结合要求为分析建立了异常高的特异性,消除了大多数非特异性抗体相互作用。在这项研究中,单层杂交莱姆病ELISA被证明对STTT和MTTT具有更高的敏感性,但具有相同的特异性。此外,考虑到Hybrid Lyme ELISA对迁移性红斑患者的灵敏度为> ~ 90%,该检测不仅可能将血清学检测从两步检测转变为单步检测,而且可能首次为迁移性红斑患者的诊断提供一种手段。莱姆病是一种由伯氏疏螺旋体(Borrelia burgdorferi)引起的蜱传螺旋体感染,其诊断存在两大局限性:需要两层血清学检测算法以提供足够的特异性;该算法在实践中对早期莱姆病的检测灵敏度较低,早期莱姆病表现为最常见的临床表现:皮肤移行性红斑。这项研究首次描述了一种新的检测方法,即混合莱姆病ELISA,它具有足够高的灵敏度,可以潜在地诊断超过90%的移动性红斑患者,并且具有足够高的特异性,可以排除二级检测的需要。这些试验特点表明,混合莱姆病ELISA有可能成为适用于莱姆病所有阶段实验室诊断的第一个单层血清学试验。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Journal of Clinical Microbiology
Journal of Clinical Microbiology 医学-微生物学
CiteScore
17.10
自引率
4.30%
发文量
347
审稿时长
3 months
期刊介绍: The Journal of Clinical Microbiology® disseminates the latest research concerning the laboratory diagnosis of human and animal infections, along with the laboratory's role in epidemiology and the management of infectious diseases.
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