Respiratory and intestinal diseases in pigs can have significant negative influence on productivity and animal welfare. A wide range of real-time PCR (rtPCR) assays are used in our laboratory (National Veterinary Institute, Technical University of Denmark) for pathogen detection, and PCR analyses are performed on traditional rtPCR platforms in which a limited number of samples can be analyzed per day given limitations in equipment and personnel. To mitigate these restrictions, rtPCR assays have been optimized for the high-throughput rtPCR BioMark platform (Fluidigm). Using this platform, we developed a high-throughput detection system that can be used for simultaneous examination of 48 samples with detection specificity for 18 selected respiratory and enteric viral and bacterial pathogens of high importance to Danish pig production. The rtPCR assays were validated and optimized to run under the same reaction conditions using a BioMark 48.48 dynamic array (DA) integrated fluidic circuit chip, and the sensitivity and specificity were assessed by testing known positive samples. Performance of the 48.48DA was similar to traditional rtPCR analysis, and the specificity of the 48.48DA was high. Application of the high-throughput platform has resulted in a significant reduction in cost and working hours and has provided production herds with a new innovative service with the potential to facilitate the optimal choice of disease control strategies such as vaccination and treatment.

中文翻译：

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开发高通量实时PCR系统以检测猪中的病原体。

猪的呼吸道和肠道疾病会对生产力和动物福利产生重大负面影响。在我们的实验室（丹麦技术大学，国家兽医学院）中，广泛的实时PCR（rtPCR）分析用于病原体检测，PCR分析在传统的rtPCR平台上进行，在该平台上可以分析数量有限的样品每天在设备和人员方面受到限制。为了减轻这些限制，已经针对高通量rtPCR BioMark平台（Fluidigm）优化了rtPCR分析。使用此平台，我们开发了一种高通量检测系统，该系统可用于同时检查48个样品，对18种选定的呼吸道和肠道病毒及细菌病原体具有检测特异性，这对丹麦猪的生产非常重要。使用BioMark 48.48动态阵列（DA）集成流体电路芯片对rtPCR测定法进行了验证和优化，使其可以在相同的反应条件下运行，并且通过测试已知的阳性样品来评估灵敏度和特异性。48.48DA的性能类似于传统的rtPCR分析，并且48.48DA的特异性很高。高通量平台的应用已大大降低了成本和工作时间，并为畜群提供了一项新的创新服务，有可能促进疾病预防策略（如疫苗接种和治疗）的最佳选择。通过检测已知的阳性样品评估敏感性和特异性。48.48DA的性能类似于传统的rtPCR分析，并且48.48DA的特异性很高。高通量平台的应用已大大降低了成本和工作时间，并为畜群提供了一项新的创新服务，有可能促进疾病预防策略（如疫苗接种和治疗）的最佳选择。通过检测已知阳性样品评估敏感性和特异性。48.48DA的性能类似于传统的rtPCR分析，并且48.48DA的特异性很高。高通量平台的应用已大大降低了成本和工作时间，并为畜群提供了一项新的创新服务，有可能促进疾病预防策略（如疫苗接种和治疗）的最佳选择。