An allele-specific PCR assay for the rapid and serotype-specific detection of salmonella pullorum
Desai, A.R.; Shah, D.H.; Shringi, Smriti; Lee, M.J.; Li, Y.H.; Cho, M.R.; Park, J.H.; Eo, S.K.; Lee, J.H. and Chae, J.S. (2005) An allele-specific PCR assay for the rapid and serotype-specific detection of salmonella pullorum. Avian Diseases, 49 (4). pp. 558-564.
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Salmonella serovar Pullorum is a causative agent of pullorum disease (PD) in poultry and is responsible for severe economic losses to the poultry industry in many parts of the world. A definitive detection of Pullorum requires culture followed by serotyping and biochemical identification, a process that is tedious and takes several weeks to accomplish. We have developed a rapid allele-specific polymerase chain reaction (PCR) method based on the nucleotide polymorphism in r f bS gene sequence for the serotype-specific detection of Pullorum and its differentiation from the closely related Gallinarum. The specificity of this PCR assay was tested using DNA samples from Pullorum (n=13), Salmonella serotypes other than Pullorum (n=19), and closely related non-Salmonella organisms (n=5). The PCR assay was highly serotype-specific as the PCR amplicon of 147 base pairs was observed only in the case of Pullorum, while all the other DNA samples tested PCR negative. A definitive identification of Pullorum cultures was possible in less than 3 hr. As little as 100 pg of SP DNA was detected. This allele-specific PCR method is highly specific as well as sensitive and may be an effective molecular tool in the rapid and serotype-specific detection of Pullorum and differentiation from other Salmonella species.
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