中国兽医科学2025，Vol.55Issue(9)：1190-1197,8.DOI:10.16656/j.issn.1673-4696.2025.0181
马鼻疽和马鼻肺炎双重荧光定量PCR检测方法的建立
Dual fluorescence quantitative PCR for the detection and differentiation of glanders and equine rhinopneumonitis
摘要
Abstract
To rapidly detect the pathogenic nucleic acids of glanders and equine rhinopneumonitis in samples of imported or exported horses,two conserved gene fragments of the pathogens were selected.Suitable fluorescence quantitative PCR primers and probes were designed respectively.The optimal reaction concentrations and reaction conditions of the experiment were screened and optimized,and a dual fluorescence quantitative PCR(FQ-PCR)method for the detection of pathogenic nucleic acids of glanders and equine rhinopneumonitis was established.The results of the study revealed that the established method and system could well distinguish the pathogen nucleic acids of glanders,equine rhinopneumonitis and other equine diseases.The correlation coefficients of the dual quantitative PCR standard curves for detecting glanders and equine rhinopneumonitis were R2=0.996 7 and R2=0.996 5 respectively.The minimum detection limits for DNA content were 44.7 copies/μL and 32.6 copies/μL respectively.The results showed that the proposed method has high specificity,sensitivity and repeatability.It can be used for the rapid detection of equine diseases in laboratories and can simultaneously detect the nucleic acids of two pathogens,glanders and equine rhinopneumonitis,which has important practical application value.关键词
马鼻疽/马鼻肺炎/双重荧光定量PCRKey words
glanders/equine rhinopneumonitis/dual fluorescence quantitative PCR分类
农业科技引用本文复制引用
鱼海琼,佟铁铸,魏俊,张璐,孙铭飞,苏飞雄,李娟,梁佳琪,李明,王国胜,王莹,陈芳,吴晓薇,曾潇..马鼻疽和马鼻肺炎双重荧光定量PCR检测方法的建立[J].中国兽医科学,2025,55(9):1190-1197,8.基金项目
农业农村部禽流感等家禽重大疫病防控重点实验室/广东省畜禽疫病防治研究重点实验室开放课题(YDWS202401) （YDWS202401）
海关总署科研项目(2024HK253,2025HK048) （2024HK253,2025HK048）
