An allele-specific PCR assay for the rapid and serotype-specific detection of Salmonella pullorum |
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| Authors: | Desai Atul R Shah Devendra H Shringi Smriti Lee Mi-Jin Li Ying-Hua Cho Mae-Rim Park Jin-Ho Eo Seong-Kug Lee John-Hwa Chae Joon-Seok |
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| Institution: | Bio-Safety Research Institute, Department of Veterinary Internal Medicine and Pathology, College of Veterinary Medicine, Chonbuk National University, Jeonju, Jeonbuk, South Korea. |
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| Abstract: | 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 rfbS 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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