1.Detection of Streptococcus dysgalactiae subsp. equisimilis in equine nasopharyngeal swabs by PCR.
Silvia PREZIUSO ; Fulvio LAUS ; Aurora Romero TEJEDA ; Carlo VALENTE ; Vincenzo CUTERI
Journal of Veterinary Science 2010;11(1):67-72
Streptococcus (S.) dysgalactiae subsp. equisimilis is responsible for severe diseases in humans, including primary bacteraemia, pneumonia, endocarditis, and toxic shock syndrome. Infection in some animal species can also occur, although a few studies have looked into cross-species infectivity. In horses, S. equisimilis is generally considered infrequent or opportunistic, but has recently been isolated from cases of strangles-like disease. Rapid and sensitive diagnostic techniques could enable epidemiological studies and effective investigation of outbreaks involving these bacteria. In this study, PCR protocols previously described in cattle and in humans to detect the species S. dysgalactiae and the subspecies equisimilis were evaluated to detect specific sequences in equine samples. For this purpose, 99 monolateral nasal swabs were collected from horses from stud farms with a history of S. equisimilis infection and were tested blindly by bacteriological isolation and by single and duplex PCR. DNA for PCR was extracted both from the colonies grown on agar media and from enrichment broth aliquots after incubation with nasal swab samples. S. equisimilis was identified by bacteriological isolation in 23 out of 99 swab samples, and PCR assays on these colonies were fully concordant with bacteriological identification (kappa statistic = 1.00). In addition, PCR of the enrichment broth aliquots confirmed the bacteriological results and detected S. equisimilis in 6 samples more than the bacteriological examination (kappa statistic = 0.84). The PCR protocols appeared to be reliable for the rapid identification of S. equisimilis in equine nasal swab samples, and could be useful for microbiological diagnosis.
Animals
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NA, Bacterial/chemistry/genetics
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Horse Diseases/diagnosis/*microbiology
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Horses
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Limit of Detection
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Male
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Nasopharynx/microbiology
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Polymerase Chain Reaction/methods/*veterinary
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Respiratory Tract Infections/diagnosis/microbiology/*veterinary
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Sensitivity and Specificity
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Streptococcal Infections/diagnosis/microbiology/*veterinary
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Streptococcus/genetics/*isolation & purification
2.Development of a multiplex PCR assay to detect Edwardsiella tarda, Streptococcus parauberis, and Streptococcus iniae in olive flounder (Paralichthys olivaceus).
Seong Bin PARK ; Kyoung KWON ; In Seok CHA ; Ho Bin JANG ; Seong Won NHO ; Fernand F FAGUTAO ; Young Kyu KIM ; Jong Earn YU ; Tae Sung JUNG
Journal of Veterinary Science 2014;15(1):163-166
A multiplex PCR protocol was established to simultaneously detect major bacterial pathogens in olive flounder (Paralichthys olivaceus) including Edwardsiella (E.) tarda, Streptococcus (S.) parauberis, and S. iniae. The PCR assay was able to detect 0.01 ng of E. tarda, 0.1 ng of S. parauberis, and 1 ng of S. iniae genomic DNA. Furthermore, this technique was found to have high specificity when tested with related bacterial species. This method represents a cheaper, faster, and reliable alternative for identifying major bacterial pathogens in olive flounder, the most important farmed fish in Korea.
Animals
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Edwardsiella tarda/genetics/*isolation & purification
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Enterobacteriaceae Infections/diagnosis/microbiology/*veterinary
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Fish Diseases/*diagnosis/microbiology
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Fisheries/*methods
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*Flatfishes
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Multiplex Polymerase Chain Reaction/economics/*veterinary
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Sensitivity and Specificity
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Streptococcal Infections/diagnosis/microbiology/*veterinary
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Streptococcus/genetics/*isolation & purification