Department of Veterinary Medicine, College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu, China.
Veterinary Diagnostic Laboratory, Department of Clinical Science, College of Veterinary Medicine, University of Illinois Urbana-Champaign, Urbana, IL, USA.
J Vet Diagn Invest. 2022 Mar;34(2):302-305. doi: 10.1177/10406387211068447. Epub 2022 Feb 10.
We developed a rapid insulated isothermal PCR (iiPCR) assay for on-site detection of using a primer and probe set targeting the superoxide dismutase (A) gene. Our iiPCR assay detected clinical isolates successfully and produced negative results on other bovine or ovine respiratory pathogens, including , , , , and spp., indicating that the PCR reactions were specific. Additionally, our iiPCR assay detected as few as 21 copies of genomic DNA and 17.2 cfu/mL of bacterial culture, which was 10 and 100 times more sensitive than conventional PCR, respectively. Our iiPCR assay can be performed on a portable device in a total of 58 min and may be a useful tool for the detection of in bovine and ovine respiratory disease in the field.
我们开发了一种快速隔热等温 PCR（iiPCR）检测方法，用于现场检测 ，该方法使用针对超氧化物歧化酶（A）基因的引物和探针组。我们的 iiPCR 检测方法成功地检测到了 临床分离株，并且对其他牛或羊呼吸道病原体（包括 、 、 、 、 和 spp.）产生阴性结果，表明 PCR 反应具有特异性。此外，我们的 iiPCR 检测方法可以检测到少至 21 个基因组 DNA 拷贝和 17.2 cfu/mL 的细菌培养物，分别比传统 PCR 方法灵敏 10 倍和 100 倍。我们的 iiPCR 检测方法可以在总共 58 分钟内在便携式设备上进行，并且可能是在现场检测牛和羊呼吸道疾病中 的有用工具。
