An allele-specific PCR assay for the rapid and serotype-specific detection of Salmonella pullorum.
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.
Desai
Atul R
AR
Bio-Safety Research Institute, Department of Veterinary Internal Medicine and Pathology, College of Veterinary Medicine, Chonbuk National University, Jeonju, Jeonbuk, South Korea.
Shah
Devendra H
DH
Shringi
Smriti
S
Lee
Mi-Jin
MJ
Li
Ying-Hua
YH
Cho
Mae-Rim
MR
Park
Jin-Ho
JH
Eo
Seong-Kug
SK
Lee
John-Hwa
JH
Chae
Joon-Seok
JS
D000483:Alleles; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D005798:Genes, Bacterial; D016133:Polymerase Chain Reaction / Q000379:methods / Q000706:statistics & numerical data / Q000662:veterinary*; D011200:Poultry / Q000382:microbiology*; D011201:Poultry Diseases / Q000175:diagnosis / Q000382:microbiology; D012475:Salmonella / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D012481:Salmonella Infections, Animal / Q000175:diagnosis / Q000382:microbiology; D012680:Sensitivity and Specificity; D012703:Serotyping
D017931:DNA Primers; D004269:DNA, Bacterial