Detection and identification of salmonellas from poultry-related samples by PCR |
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| Authors: | Oliveira S D Santos L R Schuch D M T Silva A B Salle C T P Canal C W |
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| Institution: | 1. Faculdade de Veterinária, Centro de Diagnóstico e Pesquisa em Patologia Aviária (CDPA), Universidade Federal do Rio Grande do Sul (UFRGS), Avenida Bento Gonçalves 8824, 91540-000, Porto Alegre, RS, Brazil;2. Laboratório de Referência Animal (LARA), Ministério da Agricultura, Pecuária e Abastecimento (MAPA), Porto Alegre, RS, Brazil;1. Department of Dermatology, Preventive Medicine and Medical Social Sciences, Northwestern University Feinberg School of Medicine, Chicago, Illinois;2. Northwestern Medicine Multidisciplinary Eczema Center, Chicago, Illinois;3. Department of Dermatology, Oregon Health & Science University, Portland, Oregon;1. Laboratory of Avian Diseases, College of Veterinary Medicine, Seoul National University, Seoul 08826, Republic of Korea;2. Department of Farm Animal Medicine, College of Veterinary Medicine, Seoul National University, Seoul 08826, Republic of Korea;3. Research Institute for Veterinary Science, College of Veterinary Medicine, BK21 for Veterinary Science, Seoul 08826, Republic of Korea;4. Farm Animal Clinical Training and Research Center (FACTRC), GBST, Seoul National University, Kangwon-do 25354, Republic of Korea;1. Jiangsu Key Lab of Zoonosis/Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, China;2. College of Veterinary Medicine, Henan University of Animal Husbandry and Economy, Henan, China;3. Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture of China, Yangzhou University, Yangzhou, China;2. Alltech-University of Kentucky Nutrition Research Alliance, Lexington, Kentucky 40546 |
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| Abstract: | A polymerase chain reaction (PCR) assay was developed for the generic detection of Salmonella sp. and the identification of S. Enteritidis (SE), S. Gallinarum (SG), S. Pullorum (SP) and S. Typhimurium (ST) in material collected in the field from poultry. The specificity and sensitivity of the assay combined with Rappaport-Vassiliadis selective enrichment broth (PCR-RV) were determined, and field samples were analyzed to verify the validity of the method application. Specificity of the assay was tested using 29 SE, 11 SG, 10 ST and 10 SP strains, along with 75 strains of 28 other Salmonella serovars and 21 strains of other bacterial genera. The assay was 100% specific for Salmonella detection and ST identification. The primer pair for SE, SG and SP also detected S. Berta. PCR detection limits for Salmonella at the genus level were 2 ST, 8 SE, 1.1x10(3) SG and 1.8x10(5) SP cells. At the serovar level, detection limits were 7 ST, 1.2x10(3) SE, 4.4x10(7) SG and 1.8x10(6) SP cells. At the genus level, PCR-RV detected approximately 128% more positive field samples than the standard microbiological techniques and results were ready in 48h instead of 7 days. PCR-RV method is diagnostic of Salmonella at the genus level and ST at the serovar level, although other tests are needed to identify SE, SG and SP to serovar level. |
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