Abstract
Aim
We used a polymerase chain reaction (PCR) lateral flow assay1) to rapidly diagnose joint infection. We evaluated the usefulness of multiplex-PCR (PCR lateral flow assay: PCR-LF) using detailed clinical data.
Method
A total of 35 synovial fluid samples were collected from 26 patients in whom bacterial infection was suspected, including 22 from knee joints, 11 from hip joints, and 2 from other joints. After purifying the DNA from the samples, multiplex PCR targeting two MRSA-associated genes (femA and mecA) and the bacterial 16S rRNA gene was performed. Amplified gene fragments were specifically detected with DNA probes immobilized on stick devices through DNA-DNA hybridization and visualization, enabling diagnosis of MRSA, MSSA, MRCNS, gram-positive, and/or gram-negative bacterial infection. Genetic identification of bacteria by determining the 16S rRNA gene sequence was also performed using multiplex PCR-positive samples. Finally, the usefulness of our PCR-LF method was evaluated using detailed clinical data.
Results
The results of PCR-LF were 9 gram-positive and 1 gram-negative bacterial infections. Eleven bacterial species were identified based on 16S rRNA gene sequences. Ten (90.9%)of the eleven samples (bacterial species) were identified using our PCR-LF. Five samples were detected in bacterial cultures; two are MSSA, one is Streptococcus agalactiae, one is Escherichia coli, one is Prevotella oralis. We diagnosed 6 samples as clinical infections. Therefore, the sensitivity and specificity of the culture tests were 83% and 100%, respectively, while for PCR-LF, these values were 83% and 83%.
Conclusions
PCR-LF is highly sensitive and effective for the rapid diagnosis of joint infection; however, dead bacteria may also be detected. Moreover, because the target bacterial species are limited, clinical diagnosis based on the results of multiple examinations is necessary.
