Development of a Dual PCR Method for Detection of Pasteurella Multocida and Haemophilus Parasuis

Abstract


 Pasteurella multocida and Haemophilus parasuis are the main pathogens of porcine respiratory disease syndrome, and often are the common pathogens of porcine pulmonary disease in collectivized pig farms. It has brought great influence on the pig industry and seriously hampered the healthy development of pigs. Therefore, the rapid detection method for Pasteurella multocida and Haemophilus parasuis is the key to its successful prevention and control. In the process of diagnosis, we need to conduct laboratory tests for confirming of preliminary diagnosis, that can be made based on the history, clinical symptoms, pathological changes of necropsy and epidemiological characteristics of the disease. Traditional diagnostic techniques, such as bacterial isolation and immunological test, are time-consuming and laborious, and are not suitable for rapid clinical diagnosis, nor for large-scale epidemiological investigation. Therefore, it is necessary to establish an accurate and rapid method to identify the two common respiratory pathogens in pigs. 
 In this paper, Pasteurella multocida and Haemophilus parasuis were used as research objects to establish a dual PCR detection method. The main research contents are as follows: A dual PCR assay was established to detect Pasteurella multocida and Haemophilus parasuis simultaneously. By using Pasteurella multocida and Haemophilus parasuis specific primers, and adjusting primer concentration and annealing temperature, a dual PCR method for detecting Pasteurella multocida and Haemophilus parasuis was established.
 The rapid detection method for Pasteurella multocida and Haemophilus parasuis established in this study has high specificity and sensitivity, and can realize rapid and accurate identification of pathogenic bacteria in a relatively short period of time, providing a new technical means for rapid detection in clinical and grassroots laboratories