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Detection of Bacteria in Equine Synovial Fluid by Use of the Polymerase Chain Reaction
Author(s) -
CRABILL M.R.,
COHEN N.D.,
MARTIN L.J.,
SIMPSON R.B.,
BURNEY N.
Publication year - 1996
Publication title -
veterinary surgery
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.652
H-Index - 79
eISSN - 1532-950X
pISSN - 0161-3499
DOI - 10.1111/j.1532-950x.1996.tb01398.x
Subject(s) - medicine , polymerase chain reaction , bacteria , synovial fluid , microbiology and biotechnology , computational biology , pathology , genetics , gene , alternative medicine , biology , osteoarthritis
Equine synovial fluid aliquots were inoculated with Salmonella enteritidis, Escherichia coli, Actinobacillus equuli, Staphylococcus aureus , and Streptococcus zooepidemicus to obtain approximate concentrations of 1000, 100, 10, and 1 colony forming U/mL. Synovial fluid aliquots were also inoculated with an unquantitated inoculum of Bacteroides fragilis and Clostridium perfringens. Inoculated synovial fluid was incubated in trypticase‐soy broth or Columbia broth for approximately 12 hours. Then aliquots were removed for DNA extraction and polymerase chain reaction (PCR) analysis for detection of a 531 base‐pair segment of bacterial DNA corresponding to a region of the 16S ribosomal gene. Duplicate samples of inoculated synovial fluid were prepared for microbial culture. Bacteria were detected in all samples inoculated with bacteria but not in control synovial fluid samples. Under experimental conditions there was no difference between microbial culture and PCR analyses for detection of bacteria. Experimentally, PCR was able to detect bacteria in synovial fluid within 24 hours of inoculation.