Development of a one-run real-time PCR detection system for pathogens associated with porcine respiratory diseases

SUNAGA, Fujiko; TSUCHIAKA, Shinobu; KISHIMOTO, Mai; AOKI, Hiroshi; KAKINOKI, Mari; KURE, Katsumasa; OKUMURA, Hanako; OKUMURA, Maho; OKUMURA, Atsushi; NAGAI, Makoto; OMATSU, Tsutomu; MIZUTANI, Tetsuya

doi: 10 .1292/jvms. bronchiseptica, Haemophilus parasuis, Mycoplasma hyorhinis, Mycoplasma hyosynovie, pseudorabies virus (PRV), porcine respiratory corona virus (PRCV), Porcine cytomegalovirus (PCMV) [7, 8, 15, 20] . Infection with each single pathogen does not necessarily result in appearance of symptoms, but complex infections with a variety of pathogens, including the indigenous agents, develop severe conditions. Infections with such multiple pathogens make it difficult to rapidly identify the etiology of PRDC. To adopt appropriate measures, such as vaccination or hygiene management, and to minimize the economic loss of PRDC, it is necessary to quickly, accurately and comprehensively detect multiple pathogens present in varying proportions in each herd. Serological tests [13] , pathogen isolation [22] and PCR-based tests [1, 11] are currently available to diagnose PRDC in laboratories. Most tests are based on a one assay-one pathogen approach, and they are not enough for diagnosis of PRDC in terms of comprehensiveness and rapidity. Tsuchiaka et al. previously developed a system to detect microbes in bovine diarrhea by TaqMan real-time PCR, permitting the simultaneous screening of 19 pathogens associated with diarrhea [26] . TaqMan real-time PCR possesses the advantages of high sensitivity, high specificity, and simple operation.