DETECTION OF BACTERIA IN EQUINE SYNOVIAL-FLUID BY USE OF THE POLYMERASE CHAIN-REACTION

Equine synovial fluid aliquots were inoculated with Salmonella enterit idis, Escherichia coli, Actinobacillus equuli, Staphylococcus aureus, and Streptococcus zooepidemicus to obtain approximate concentrations o f 1000, 100, 10, and 1 colony forming U/mL. Synovial fluid aliquots we re also inoculated with an unquantitated inoculum of Bacteroides fragi lis and Clostridium perfringens. Inoculated synovial fluid was incubat ed in trypticase-soy broth or Columbia broth for approximately 12 hour s. Then aliquots were removed for DNA extraction and polymerase chain reaction (PCR) analysis for detection of a 531 base-pair segment of ba cterial DNA corresponding to a region of the 16S ribosomal gene. Dupli cate samples of inoculated synovial fluid were prepared for microbial culture. Bacteria were detected in all samples inoculated with bacteri a but not in control synovial fluid samples. Under experimental condit ions there was no difference between microbial culture and PCR analyse s for detection of bacteria. Experimentally, PCR was able to detect ba cteria in synovial fluid within 24 hours of inoculation. (C)Copyright 1996 by The American College of Veterinary Surgeons