关键词: <, p>, 流感嗜血杆菌, 溶血嗜血杆菌, 多杀巴斯德杆菌, 多重荧光定量 PCＲ, 实验动物<, /p>

Abstract: Abstract: Objective To develop a multiplex real-time quantitative PCＲ ( qPCＲ) assay to detect Haemophilus influenzae，Haemophilus haemolyticus and Pasteurella multocida in laboratory animals． Method We establish the multiple qPCＲ reaction system by specificity，sensitivity and reproducibility verification，according to Hpd gene of H． influenzae and H． haemolyticus，KMT1 gene of P． multocida designed specific primers and Taqman probes respectively． Then，the 822 laboratory animal respiratory samples were detecting by the assay application． Ｒesult There was no cross reaction between the three pathogens and 29 others． The sensitivity of the multiple qPCＲ method detecting above three pathogens could reach 10 copies /μL． Among the tree shrew specimens，the positive rate of H． haemolyticus and P． multocida were 10% ( 6 /60) and 21. 7% ( 13 /60) respectively，but other animals were negative in three pathogens． Conclusion The multiple qPCＲ assay in this study that can quickly detect the above three kinds of respiratory pathogen in laboratory animals．

Key words: <p>Haemophilus influenza, Haemophilus haemolyticus, Pasteurella multocida, Ｒeal time PCＲ, Laboratory animals</p>