LeCuyer Tessa E, Franklin-Guild Rebecca, Guarino Cassandra, Fox Alexandra, Maddock Kelli, Barber Rebecca, Baum David H, Bustamante Felipe, Daniels Joshua, de Avila David M, Diaz-Campos Dubraska, Glick Lisa G, Haley Jessica, Heinen Sheila, Leger Laura, Loy John Dustin, Pillai Deepti, Poonsuk Korakrit, Russell Michael M, Shukla Mithila, Schwarz Erika R, Stempien Matthew, Tewari Deepanker, van Balen Joany C, Werre Stephen R, Cecere Julie T
Department of Pathology, Microbiology, and Immunology, School of Veterinary Medicine, University of California, Davis, Davis, CA, United States.
Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, United States.
Front Vet Sci. 2025 Apr 1;12:1556965. doi: 10.3389/fvets.2025.1556965. eCollection 2025.
is a zoonotic pathogen of dogs that poses diagnostic challenges. While direct detection of by PCR or culture is ideal, serologic diagnosis is necessary for identification of carrier animals and can support a clinical diagnosis of brucellosis. Prior to 2022, seroscreening in the United States was primarily performed using a commercially available rapid slide agglutination test. However, the kit was discontinued by the manufacturer in early 2022, leaving a gap in the availability of commercial seroassays. The goal of this study was to compare the performance of three serologic tests that are currently available: VMRD ELISA, Bionote Anigen Rapid C.Brucella Ab immunochromatographic lateral flow assay, and VMRD indirect fluorescent antibody (IFA) assay. A panel of 56 banked serum specimens originally submitted to the Cornell University Animal Health Diagnostic Center (the study reference laboratory) for seroscreening was distributed to 12 testing laboratories. Each sample was run on three assays developed at the reference lab: rapid slide agglutination test with 2-mercaptoethanol (2-ME RSAT), agar gel immunodiffusion test using cytoplasmic antigen (AGID II), and Canine Brucella Multiplex. Five testing labs ran the ELISA, six ran the lateral flow, and six ran the IFA. When evaluated as a screening assay, we compared the assays to the 2ME-RSAT. The ELISA had the highest sensitivity (96.8, 95%CI 83.8-99.9) but the lowest specificity (79.3, 95%CI 57.9-92.9). The sensitivity of the lateral flow was 90.6% (95%CI 75-98%) and the IFA was 87.5% (95%CI 71-96.5). Specificity for the lateral flow was 95.8% (95%CI 78.9-99.9) and IFA was 97.5% (95%CI 67.6-97.3). When compared to AGID II and Canine Brucella Multiplex, the test assays were all highly sensitive, but specificity was <90%. Interrater reliability was highest for IFA ( = 0.92) and lowest for the lateral flow ( = 0.82). Serial testing of positive samples with a more specific test, such as AGID II, will continue to be necessary when using any of the three assays tested in this study.
是一种给犬类诊断带来挑战的人畜共患病原体。虽然通过聚合酶链反应（PCR）或培养直接检测是理想的，但血清学诊断对于识别携带动物是必要的，并且可以支持布鲁氏菌病的临床诊断。在2022年之前，美国的血清学筛查主要使用市售的快速玻片凝集试验。然而，该试剂盒在2022年初被制造商停产，导致商业性布鲁氏菌血清学检测方法出现空白。本研究的目的是比较目前可用的三种布鲁氏菌血清学检测方法的性能：VMRD酶联免疫吸附测定（ELISA）、Bionote Anigen Rapid C.Brucella Ab免疫层析侧向流动测定和VMRD间接荧光抗体（IFA）测定。一组最初提交给康奈尔大学动物健康诊断中心（该研究的参考实验室）进行布鲁氏菌血清学筛查的56份储存血清标本被分发给12个检测实验室。每个样本在参考实验室开发的三种检测方法上进行检测：使用2-巯基乙醇的快速玻片凝集试验（2-ME RSAT）、使用细胞质抗原的琼脂凝胶免疫扩散试验（AGID II）和犬布鲁氏菌多重检测。五个检测实验室进行ELISA检测，六个进行侧向流动检测，六个进行IFA检测。当作为筛查检测进行评估时，我们将这些检测方法与2ME-RSAT进行比较。ELISA具有最高的灵敏度（96.8，95%置信区间83.8-99.9），但特异性最低（79.3，95%置信区间57.9-92.9）。侧向流动检测的灵敏度为90.6%（95%置信区间75-98%），IFA为87.5%（95%置信区间71-96.5）。侧向流动检测的特异性为95.8%（95%置信区间78.9-99.9），IFA为97.5%（95%置信区间67.6-97.3）。与AGID II和犬布鲁氏菌多重检测相比，这些检测方法都具有高度敏感性，但特异性<90%。评分者间可靠性IFA最高（κ=0.92），侧向流动检测最低（κ=0.82）。当使用本研究中测试的三种检测方法中的任何一种时，继续使用更特异的检测方法（如AGID II）对阳性样本进行系列检测将仍然是必要的。
