Development of an improved species specific PCR test for detection of Haemophilus parasuis.

摘要: A PCR test for identification of Haemophilus parasuis was optimized using the 16S rDNA sequences 15 serotype reference strains H. parasuis. The evaluated on a collection 218 Danish field isolates as well 81 representatives 27 other species, including genetically affiliated species within Pasteurellaceae. In addition, DNA preparations from 56 North America were included. To obtain that specific parasuis, multiplex 3 different primers developed. produced an amplicon approximately 1090 bp only with further 55 clinical samples 16 pigs suspected being infected showing polyserositis or septicemia at autopsy 492 nasal swabs. compared performance earlier published by Oliveira et al. [Oliveira, S., Galina, L., Pijoan, C., 2001. Development to diagnose infections. J. Vet. Diagn. Invest. 13, 495-501]. sensitivity present found be slightly lower when applied diseased and 10-fold tested pure cultures (5CFU 0.5CFU/PCR reaction, respectively). Addition 1.4 x 10(5) Escherichia coli each tube did not alter tests. No difference in tests observed purified DNA. On hand, 100% contrast al., which also positive belonging A. indolicus, porcinus, minor, commonly occurring upper respiratory tract. However, is used systemic locations chances false results are apparently low. represents rapid reliable method based high specificity makes it suitable detection samples, regardless presence contaminating flora. As two differ specificity, use both purposes possibility.