Overview

Synovial fluid is secreted into the joint cavity from the inner membrane of synovial joints. Synovial fluid is a plasma ultrafiltrate which contains proteins derived from both the blood plasma and produced by cells within the joint tissues. It lubricates the articulating joints as well as supplies oxygen and nutrients while removing carbon dioxide and metabolic wastes from the chondrocytes in the surrounding cartilage. Septic arthritis is usually caused by bacterial, viral or fungal infections to the synovial fluid. Staphylococcus aureus is the most common bacterial infection causing Septic arthritis. Diagnosing Septic arthritis is done through joint fluid (synovial fluid) analysis, blood tests or imaging tests.  

Norgen’s Synovial Fluid Bacterial Genomic DNA Isolation Kit provides a fast, reliable and simple procedure for isolating the highest quality and the highest quantity of bacterial genomic DNA from various amounts of synovial fluid ranging from 1 mL up to 10 mL.  Purification is based on using Norgen’s proprietary resin separation matrix. The kit is designed to isolate all Gram +ve and Gram -ve strains. Moreover, the kit allows the user to elute the purified bacterial genomic DNA into a flexible elution volume ranging from 25 µL to 50 µL. The purified bacterial gDNA is eluted in an Elution Buffer that is compatible with all downstream applications including PCR, qPCR, methylation-sensitive PCR, Southern Blot analysis and NGS.

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Supporting Data

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Figure 1.  Isolation and Detection of the Gram +Ve S. Aureus Bacterial Genomic DNA from 1 mL of spike-in Synovial fluid. Genomic DNA was isolated from 1 mL synovial fluid spiked with a serially diluted Gram +Ve S. Aureus using Norgen’s Synovial Fluid Bacterial genomic DNA Purification Kit. The efficiency of the purified bacterial gDNA from the spiked-in synovial fluid was evaluated against the gDNA isolation from pure culture containing the same amount of S. Aureus spiked in 1mL synovial fluid. The purified gDNA was subsequently detected using quantitative PCR.  All serially diluted spiked-in Gram +Ve S. Aureus was purified with high efficiency from 1mL  synovial fluid as compared to the amplification of S. Aureus from pure culture. The limit of detection for the Gram +Ve S. Aureus was down to 5 CFU/mL.   

Figure 2.  Isolation and Detection of the Gram -Ve E.coli Bacterial Genomic DNA from 1 mL of spike-in Synovial fluid. Genomic DNA was isolated from 1 mL synovial fluid spiked with a serially diluted Gram -Ve E.coli using Norgen’s Synovial Fluid Bacterial genomic DNA Purification Kit. The efficiency of the purified bacterial gDNA from the spiked-in synovial fluid was evaluated against the gDNA isolation from pure culture containing the same amount of E.coli spiked in 1mL synovial fluid. The purified gDNA was subsequently detected using quantitative PCR.  All serially diluted spiked-in Gram -Ve E.coli was purified with high efficiency from 1mL  synovial fluid as compared to the amplification of E.coli from pure culture. The limit of detection for the Gram -Ve E.coli was down to 5 CFU/mL.  

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