Detection of bacteria in equine synovial fluid by use of the polymerase chain reaction.

The research focuses on detecting bacteria in horse’s synovial fluid using Polymerase Chain Reaction. The researchers exposed the synovial fluid to various types of bacteria and discovered that they could identify the presence of these bacteria in less than 24 hours using PCR.

Research Process

• The research team started by obtaining equine synovial fluid aliquots (samples of synovial fluid from horses). This type of fluid is found in the cavities of synovial joints such as the knee, shoulder, and hip.
• These aliquots were exposed to various types of bacteria, namely, Salmonella enteritidis, Escherichia coli, Actinobacillus equuli, Staphylococcus aureus, and Streptococcus zooepidemicus. The aim was to have different concentrations of 1000, 100, 10, and 1 colony forming unit per mL.
• Additionally, each aliquot was also exposed to an undefined amount of Bacteroides fragilis and Clostridium perfringens.
• After inoculation, the synovial fluid was incubated in either a trypticase-soy broth or Columbia broth for roughly 12 hours.

Analysis and Detection

• After incubation, samples were taken from each aliquot for DNA extraction and Polymerase Chain Reaction (PCR) analysis.
• The PCR process targeted a specific 531 base-pair segment of bacterial DNA that corresponds with an area of the 16S ribosomal gene, known for its use in identifying and classifying bacteria.
• Duplicate samples were also prepared for microbial culture, to compare the effectiveness between traditional microbial culture and the PCR analysis.
• In all cases, bacteria were detected in the samples that had been inoculated with bacteria, but not in the control samples (samples of synovial fluid not exposed to bacteria).
• The results of the PCR analyses were comparable to those of traditional microbial culture methods. Importantly, PCR analysis was able to detect bacteria presence in synovial fluid within 24 hours of inoculation, providing a fast method for bacterial detection.

Conclusions

• The research concluded that PCR is an effective tool for detecting specific bacterial DNA in equine synovial fluid under experimental conditions.
• PCR also offers some advantages over traditional microbial culture techniques; notably, the time required for detecting bacteria is significantly reduced, with results obtained within 24 hours post-inoculation.