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Updated: Nov 9, 2025
Identification of Rare Bacterial Pathogens by 16S rRNA Gene Sequencing and MALDI-TOF MS
Published on: July 11, 2016
Efficacy of 16S rRNA variable regions high-resolution melt analysis for bacterial pathogens identification in
Samaneh Bourbour1, Mohammad Emaneini1, Mahmoud Jabalameli2
1Department of Microbiology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Background:
Accurate and rapid identification of microorganisms causing periprosthetic joint infections (PJIs) are necessary for choosing an appropriate antibiotic therapy. Therefore, molecular techniques are suggested for diagnosis in suspected PJIs. The Broad-range PCR and High-Resolution Melt Analysis (HRMA) were evaluated for the identification of causative organisms of PJIs in this study.
Results:
For 47 of 63 specimens, both the culture and broad-range PCR were positive. The culture was found to be able of organism's detection in 74.6% (47/63) of patients. Of 47 positive cultures, 11 (23.4%) were polymicrobial and 36 (76.59%) were monomicrobial cultures, in which 34 (91.89%) cases were detected by HRM assay. The sensitivity, specificity of HRMA vs monomicrobial culture were 91.89, 93.75%, respectively. The sensitivity, specificity of total HRMA (mono + poly) vs culture were 82.92, 93.75%.
Conclusions:
HRM assay coupled with broad-range PCR are effective screening, rapid, and relatively cost-effective methods for discrimination of PJIs especially in aiding culture method. Using computer programs such as the Matlab-2018b program for HRM data analysis is also valuable and helpful in diagnosis.
Insights
Broad-range PCR and High-Resolution Melt Analysis (HRMA) effectively identify causative organisms in periprosthetic joint infections (PJIs). This molecular approach aids in rapid diagnosis and guides appropriate antibiotic therapy for improved patient outcomes.
Area of Science:
- Medical Microbiology
- Molecular Diagnostics
- Infectious Diseases
Background:
- Periprosthetic joint infections (PJIs) require accurate and rapid microbial identification for effective antibiotic treatment.
- Molecular diagnostic techniques are increasingly recommended for suspected PJIs.
- Broad-range PCR and High-Resolution Melt Analysis (HRMA) are emerging molecular tools for pathogen detection.
Purpose of the Study:
- To evaluate the efficacy of broad-range PCR combined with HRMA for identifying microorganisms in PJIs.
- To compare the performance of molecular methods against traditional culture techniques.
- To assess the utility of HRMA in differentiating monomicrobial and polymicrobial infections.
Main Methods:
- Broad-range PCR was performed on 63 specimens from patients with suspected PJIs.
- High-Resolution Melt Analysis (HRMA) was utilized for microbial identification following PCR.
- Results were compared with conventional culture methods for sensitivity and specificity calculations.
Main Results:
- Broad-range PCR and culture were positive in 47 out of 63 specimens (74.6%).
- HRM assay successfully identified pathogens in 34 out of 36 monomicrobial cultures (91.89%).
- The sensitivity and specificity of HRMA compared to monomicrobial cultures were 91.89% and 93.75%, respectively.
Conclusions:
- Broad-range PCR coupled with HRMA offers a rapid, cost-effective screening method for PJI diagnosis.
- These molecular techniques effectively aid traditional culture methods in identifying causative pathogens.
- Utilizing computational tools like Matlab-2018b enhances HRM data analysis for improved diagnostic accuracy.
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