Differential identification of Salmonella enterica subsp. enterica serovar Gallinarum biovars Gallinarum and Pullorum based on polymorphic regions of glgC and speC genes.
Salmonella enterica subsp. enterica serovar Gallinarum biovars Gallinarum and Pullorum cause fowl typhoid and pullorum disease in avian species, respectively, and have been of considerable economic importance to the poultry industry in parts of the world. The definitive diagnosis of these diseases can be made only by isolation and identification of the causative agent. However, rapid identification of biovars Gallinarum and Pullorum is not easily feasible due to their common antigenic structure and genomic sequence similarity. We developed a duplex polymerase chain reaction (PCR) assay to identify and discriminate between strains of biovars Gallinarum and Pullorum. Duplex PCR primers were designed to target polymorphic regions of glgC and speC genes showing multiple mutations in the sequenced S. enterica subsp. enterica serovar Gallinarum 287/91 genome and were applied to the specific identification of biovars Gallinarum and Pullorum. Boiled lysates of 131 reference and field strains of Salmonella and other related Gram-negative bacteria were tested to validate the duplex PCR assay. All strains of biovars Gallinarum (n=53) and Pullorum (n=21) tested were correctly identified based on this assay (100% sensitivity) while the other strains (n=57) were PCR negative (100% specificity). These results demonstrate that a highly accurate biovar-specific duplex PCR assay can be performed for the rapid identification and discrimination of biovars Gallinarum and Pullorum from field isolates.
National Veterinary Research & Quarantine Service, Ministry for Food, Agriculture, Forestry and Fisheries, Anyang, Gyeonggi, Republic of Korea.
Min-Su
MS
Kang
Yong-Kuk
YK
Kwon
Byeong-Yeal
BY
Jung
Aeran
A
Kim
Kang-Mu
KM
Lee
Byung-Ki
BK
An
Eun-A
EA
Song
Jun-Hun
JH
Kwon
Gab-Soo
GS
Chung
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D001715:Bird Diseases / Q000175:diagnosis* / Q000382:microbiology; D001717:Birds; D005098:Exotoxins / Q000235:genetics*; D005798:Genes, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D012481:Salmonella Infections, Animal / Q000175:diagnosis* / Q000382:microbiology; D019779:Salmonella enterica / Q000145:classification / Q000235:genetics*; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D013045:Species Specificity
D001426:Bacterial Proteins; D005098:Exotoxins; C027951:erythrogenic toxin