Polymerase Chain Reaction Assay Using the Restriction Fragment Length Polymorphism Technique in the Detection of
Ataollah Moshirabadi1, Mohammad Razi2, Peyman Arasteh3
1Department of Orthopedic Surgery, Atieh Private Hospital, Tehran, Iran.
Background:
PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) techniques have been used for the diagnosis of bacteria in some infections. In this study, we aimed to evaluate the diagnostic accuracy of PCR for the diagnosis of prosthetic joint infections (PJI) and to identify isolated microorganisms, using the RFLP method.
Methods:
During January 2015 to January 2018, patients who were suspected of having PJI after arthroplasty surgery or were candidates for revision surgery due to loosening of implant entered the study. Patients who had 1 major criterion or 3 minor criteria for PJI based on the Philadelphia Consensus Criteria (PCC) on Periprosthetic Joint Infection were considered as cases of PJI. Both culture results and PCR findings, were cross compared with results of the PCC (as the gold standard criteria).
Results:
Overall, 76 samples were included in the study. Mean (standard deviation) age of patients was 66.72 ± 11.82 years. Overall, 57.9% of patients were females. Prevalence of PJI was 50% based on the PCC. Sensitivity, specificity, positive predictive value, negative predictive value, and general efficacy of PCR for detection of PJI was 97.4%, 100%, 100%, 97.4%, and 98.7%, respectively. Sensitivity, specificity, positive predictive value, negative predictive value, and general efficacy of culture was 31.6%, 100%, 65.7%, 100%, and 59.4%, respectively. We isolated a broad range of bacteria using PCR-RFLP including Gram-positive cocci such as Staphylococcus sp., Streptococcus sp., and Enterococcus sp., and Gram-negative bacilli such as Enterobacteriaceae sp., Pseudomonas sp. Citrobacter sp., as well as Chlamydophila pneumonia, Stenotrophomonas maltophilia, Brucella melitensis, non-gonococcal Neisseria, Kingella kingae, Bacteroides ovatus, and Proteus mirabilis from PJI patients.
Conclusion:
Inhere, for the first time, we showed that PCR-RFLP is a powerful tool for identifying the type of bacteria involved in PJI, and can be used for follow-up of patients suspected of PJI and those with a history of antibiotic use. PCR-RFLP may be able to substantially decrease detection time of PJI among PCR-based methods, while allowing more accurate identification of the bacteria involved.
Insights
Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) accurately diagnoses prosthetic joint infections (PJI), identifying diverse bacteria. This method offers faster and more precise detection than traditional cultures for PJI diagnosis.
Area of Science:
- Microbiology
- Infectious Diseases
- Molecular Diagnostics
Background:
- Prosthetic joint infections (PJI) pose significant challenges in orthopedic surgery.
- Accurate and rapid diagnosis of PJI is crucial for effective patient management and treatment.
- Current diagnostic methods, including microbial cultures, may have limitations in sensitivity and speed.
Purpose of the Study:
- To evaluate the diagnostic accuracy of polymerase chain reaction (PCR) for PJI detection.
- To identify microorganisms involved in PJI using the PCR-restriction fragment length polymorphism (PCR-RFLP) technique.
- To compare the efficacy of PCR-RFLP with traditional culture methods for PJI diagnosis.
Main Methods:
- The study included patients suspected of PJI based on Philadelphia Consensus Criteria (PCC).
- PCR and microbial cultures were performed on patient samples.
- PCR-RFLP was employed for bacterial identification, with results compared against PCC as the gold standard.
Main Results:
- PCR demonstrated high diagnostic accuracy for PJI, with sensitivity of 97.4% and specificity of 100%.
- Culture methods showed lower sensitivity (31.6%) but 100% specificity for PJI detection.
- PCR-RFLP successfully identified a wide range of Gram-positive and Gram-negative bacteria, as well as other pathogens, in PJI patients.
Conclusions:
- PCR-RFLP is a powerful tool for identifying bacterial types in PJI, outperforming traditional cultures.
- This method can significantly reduce detection time for PJI compared to other PCR-based techniques.
- PCR-RFLP aids in the follow-up of PJI patients, especially those with prior antibiotic use, enabling more accurate bacterial identification.
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