Qi Jie, Li Penghui, Yan Yasong, Li Gongmei, Kong Lingcong
College of Veterinary Medicine, Jilin Agricultural University, Changchun, China.
Open Life Sci. 2024 Mar 26;19(1):20220778. doi: 10.1515/biol-2022-0778. eCollection 2024.
Bovine respiratory disease (BRD) is a significant veterinary challenge, often exacerbated by pathogen resistance, hindering effective treatment. Traditional testing methods for primary pathogens - , , and - are notably time-consuming and lack the rapidity required for effective clinical decision-making. This study introduces a TaqMan MGB probe detection chip, utilizing fluorescent quantitative PCR, targeting key BRD pathogens and associated drug-resistant genes and sites. We developed 94 specific probes and primers, embedded into a detection chip, demonstrating notable specificity, repeatability, and sensitivity, reducing testing time to under 1 h. Additionally, we formulated probes to detect mutations in the quinolone resistance-determining region, associated with fluoroquinolone resistance in BRD pathogens. The chip exhibited robust sensitivity and specificity, enabling rapid detection of drug-resistant mutations in clinical samples. This methodology significantly expedites the diagnostic process for BRD and sensitive drug screening, presenting a practical advancement in the field.
牛呼吸道疾病（BRD）是一项重大的兽医挑战，常常因病原体耐药性而加剧，阻碍了有效治疗。针对主要病原体（ 、 和 ）的传统检测方法耗时显著，缺乏有效临床决策所需的快速性。本研究引入了一种TaqMan MGB探针检测芯片，利用荧光定量PCR，靶向关键的BRD病原体以及相关耐药基因和位点。我们开发了94种特异性探针和引物，嵌入到检测芯片中，显示出显著的特异性、重复性和灵敏度，将检测时间缩短至1小时以内。此外，我们还设计了用于检测喹诺酮耐药决定区突变的探针，这些突变与BRD病原体中的氟喹诺酮耐药性相关。该芯片展现出强大的灵敏度和特异性，能够快速检测临床样本中的耐药突变。这种方法显著加快了BRD的诊断过程和敏感药物筛选，是该领域一项切实的进展。
