Nájera-Rivera Héctor D, Rodríguez-Cortez Ana D, Anaya-Santillán María G, Díaz-Aparicio Efrén, Ramos-Rodríguez Ariadna V, Siliceo-Cantero Irlanda J, Vázquez-Franco Norma C, Nieto-Patlán Erik, Peñas Alejandro De Las, Valdés-Vázquez Liliana M, Cobos-Marín Laura
Department of Reproduction, Veterinary School, UNAM, Mexico City, Mexico.
Department of Microbiology and Immunology, Veterinary School, UNAM, Mexico City, Mexico.
Vet World. 2023 Apr;16(4):704-710. doi: 10.14202/vetworld.2023.704-710. Epub 2023 Apr 8.
Brucellosis, paratuberculosis (PTb), and infections caused by small ruminant lentivirus (SRLV), formerly known as caprine arthritis encephalitis virus (CAEV), adversely affect goat production systems. Nonetheless, commonly used diagnostic tests can only determine one analyte at a time, increasing disease surveillance costs, and limiting their routine use. This study aimed to design and validate a multiplex assay for antibody detection against these three diseases simultaneously.
Two recombinant proteins from the SRLV (p16 and gp38), the native hapten of , and the paratuberculosis-protoplasmic antigen 3 from subsp. paratuberculosis (MAP) were used to devise and assess a multiplex assay. Conditions for the Luminex multiplex test were established and validated by sensitivity, specificity, repeatability, and reproducibility parameters. Cut-off points for each antigen were also established.
The 3-plex assay had high sensitivity (84%) and specificity (95%). The maximum coefficients of variation were 23.8% and 20.5% for negative and positive control samples, respectively. The p16 and gp38 SRLV antigens are 97% and 95%, similar to the CAEV sequence found in GenBank, respectively.
The multiplex test can be effectively used for the simultaneous detection of antibodies against SRLV, MAP and in goats.
布鲁氏菌病、副结核病（PTb）以及由小反刍兽慢病毒（SRLV，原称山羊关节炎脑炎病毒，CAEV）引起的感染，对山羊生产系统产生不利影响。然而，常用的诊断检测每次只能测定一种分析物，增加了疾病监测成本，并限制了其常规应用。本研究旨在设计并验证一种可同时检测针对这三种疾病抗体的多重检测方法。
使用来自SRLV的两种重组蛋白（p16和gp38）、[此处原文缺失相关内容]的天然半抗原以及副结核亚种副结核分枝杆菌（MAP）的副结核原生质体抗原3来设计和评估多重检测方法。通过灵敏度、特异性、重复性和再现性参数建立并验证了Luminex多重检测的条件。还确定了每种抗原的临界值。
三联检测具有高灵敏度（84%）和特异性（95%）。阴性和阳性对照样品的最大变异系数分别为23.8%和20.5%。SRLV的p16和gp38抗原分别与GenBank中发现的CAEV序列相似度为97%和95%。
该多重检测方法可有效用于同时检测山羊体内针对SRLV、MAP和[此处原文缺失相关内容]的抗体。
