Laboratorio Central de Veterinaria (LCV), Ministry of Agriculture, Fisheries and Food, Ctra. M-106, Km 1,4, 28110 Algete, Spain.
Viruses. 2024 Jun 4;16(6):915. doi: 10.3390/v16060915.
The serological surveillance of bluetongue in bulk tank milk is an efficient and cost-effective method for the early detection of bluetongue virus incursions in unvaccinated free areas of the disease. In addition, the availability of standardized and reliable reagents and refined diagnostic procedures with high sensitivity and specificity are essential for surveillance purposes. However, no available reference materials for bluetongue virus serological surveillance in bulk tank milk exist. This study shows the production and characterization of reference material for the implementation of a commercially available bluetongue milk ELISA test in official laboratories, as well as the evaluation of a procedure to increase the sensitivity in samples with low levels of antibodies. This procedure, based on milk protein concentration, allowed us to notably increase the ELISA test's analytical sensitivity, which is useful for milk samples from farms with low within-herd prevalence or pools of bulk tank milk samples. The standardized milk reference material produced here, together with the evaluated procedure to improve analytical sensitivity, could be applied as tools to ensure an accurate diagnosis by official laboratories in bluetongue unvaccinated free areas.
在未接种疫苗的蓝舌病自由区，通过对大容量奶罐奶进行血清学监测，是一种早期发现蓝舌病病毒入侵的有效且具有成本效益的方法。此外，还需要有标准化和可靠的试剂以及具有高灵敏度和特异性的精细诊断程序，以满足监测目的。然而，大容量奶罐奶中蓝舌病病毒血清学监测尚无可用的参考材料。本研究展示了参考材料的生产和特性，以实施官方实验室中市售蓝舌病奶 ELISA 检测，并评估提高低抗体水平样品灵敏度的程序。该程序基于牛奶蛋白浓度，显著提高了 ELISA 检测的分析灵敏度，这对于低群体内流行率或大容量奶罐奶样池的牛奶样品非常有用。此处生产的标准化牛奶参考材料，以及评估的提高分析灵敏度的程序，可作为工具，确保未接种疫苗的蓝舌病自由区的官方实验室做出准确诊断。
