Detection and identification of avian mycoplasmas by polymerase chain reaction and restriction fragment length polymorphism assay |
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| Institution: | 1. Veterinary Medical Research Institute, Hungarian Academy of Sciences, P.O. Box 18, H-1581 Budapest, Hungary;2. Veterinary Institute of Debrecen, P.O. Box 51, H-4002 Debrecen, Hungary;3. National Veterinary Institute, P.O. Box 7073, S-750 07 Uppsala, Sweden;1. Institut national de recherche et de Sécurité, INRS, 1 rue du MORVAN, 54500 Vandoeuvre-lès-Nancy, France;2. Université de Lorraine, LEMTA, UMR 7563, 2 Avenue de la forêt de Haye, Nancy F-54000, France;1. Department of Microbiology, Nihon University School of Dentistry, Tokyo 101-8310, Japan;2. Immersion Physics Class, Department of Science, Tokyo Gakugei University International Secondary School, Tokyo 178-0063, Japan |
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| Abstract: | The polymerase chain reaction (PCR) with primers complementary to the 16S rRNA genes was used to detect avian mycoplasmas. A primer pair designed for the detection of human and rodent mycoplasmal species was examined for its ability to detect the most important avian mycoplasmas. After testing the respective reference strains, we found that Mycoplasma iowae, Mycoplasma meleagridis and Mycoplasma synoviae could be detected by PCR with this primer pair, and distinction could be made among them by restriction fragment length polymorphism (RFLP) assay with two restriction enzymes (BamHI and RsaI). For the detection of Mycoplasma gallisepticum by PCR, we needed species-specific primers. The results of the PCR- and RFLP-based identification procedures of 17 different field isolates agreed with those obtained by conventional methods. |
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