Milton A Arun Prince, Khan Sabia, Srinivas Kandhan, Basar Torik, Hussain Zakir, Priya G Bhuvana, Saminathan M, Puii L H, Ghatak Sandeep, Das Samir
Division of Animal and Fisheries Sciences, ICAR Research Complex for NEH Region, Umiam, India.
College of Agriculture, Central Agricultural University (Imphal), Kyrdemkulai, India.
Front Vet Sci. 2025 Jun 27;12:1605316. doi: 10.3389/fvets.2025.1605316. eCollection 2025.
, , and are neglected, emerging and zoonotic pathogens in pigs that share common clinical signs and lesions, particularly septicaemia and skin lesions, highlighting the need for differential diagnosis. Currently, no single diagnostic assay is available to test samples for confirmation of the presence of all three pathogens. To address this, a novel multiplex PCR-based detection assay was optimized and evaluated for the simultaneous detection of , , and targeting the, 23S rRNA, , and genes, respectively. The optimized multiplex PCR demonstrated 100% analytical specificity, with a detection limit of 10 copies/μL for and , and 10 copies/μL for . The assay was further validated by comparison with previously established uniplex PCR assays and tested on field-collected pig blood samples to assess real-world applicability. The developed assay is rapid, specific and cost-effective and can serve as a handy diagnostic tool for testing suspect samples for these neglected bacterial pathogens of pigs.
[病原体名称1]、[病原体名称2]和[病原体名称3]是猪身上被忽视的、新出现的人畜共患病原体，它们具有共同的临床症状和病变，特别是败血症和皮肤病变，这突出了鉴别诊断的必要性。目前，没有单一的诊断检测方法可用于检测样本以确认所有三种病原体的存在。为了解决这个问题，一种基于多重PCR的新型检测方法被优化并评估，用于同时检测[病原体名称1]、[病原体名称2]和[病原体名称3]，分别针对[基因名称1]、23S rRNA、[基因名称2]和[基因名称3]基因。优化后的多重PCR显示出100%的分析特异性，对[病原体名称1]和[病原体名称2]的检测限为10拷贝/μL，对[病原体名称3]的检测限为10拷贝/μL。通过与先前建立的单重PCR检测方法进行比较，进一步验证了该检测方法，并在现场采集的猪血样本上进行测试，以评估其实际应用价值。所开发的检测方法快速、特异且具有成本效益，可作为一种便捷的诊断工具，用于检测猪的这些被忽视的细菌病原体的可疑样本。
