Academic Journal
Development of a multiplex Loop-Mediated Isothermal Amplification (LAMP) for the diagnosis of bacterial periprosthetic joint infection
| العنوان: | Development of a multiplex Loop-Mediated Isothermal Amplification (LAMP) for the diagnosis of bacterial periprosthetic joint infection |
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| المؤلفون: | Jang, Woong Sik, Park, Seoyeon, Bae, Ji Hoon, Yoon, Soo Young, Lim, Chae Seung, Cho, Min-Chul |
| المساهمون: | Banada, Padmapriya P., Korea University Guro Hospital, Korea Health Industry Development Institute |
| المصدر: | PLOS ONE ; volume 19, issue 5, page e0302783 ; ISSN 1932-6203 |
| بيانات النشر: | Public Library of Science (PLoS) |
| سنة النشر: | 2024 |
| المجموعة: | PLOS Publications (via CrossRef) |
| الوصف: | Background Periprosthetic joint infection (PJI) is one of the most serious and debilitating complications that can occur after total joint arthroplasty. Therefore, early diagnosis and appropriate treatment are important for a good prognosis. Recently, molecular diagnostic methods have been widely used to detect the causative microorganisms of PJI sensitively and rapidly. The Multiplex Loop-Mediated Isothermal Amplification (LAMP) method eliminates the complex temperature cycling and delays caused by temperature transitions seen in polymerase chain reaction (PCR) methods, making it faster and easier to perform compared to PCR-based assays. Therefore, this study developed a multiplex LAMP assay for diagnosing bacterial PJI using LAMP technology and evaluated its analytical and clinical performance. Methods We developed a multiplex LAMP assay for the detection of five bacteria: Staphylococcus aureus , Staphylococcus epidermidis , Streptococcus agalactiae , Pseudomonas aeruginosa , and Escherichia coli , frequently observed to be the causative agents of PJI. The method of analytical sensitivity and cross-reactivity were determined by spiking standard strains into the joint synovial fluid. The analytical sensitivity of the multiplex LAMP assay was compared with that of a quantitative real-time PCR (qPCR) assay. Clinical performance was evaluated using 20 joint synovial fluid samples collected from patients suspected of having bacterial PJI. Results The analytical sensitivity of the gram-positive bacterial multiplex LAMP assay and qPCR were 10 5 /10 4 CFU/mL, 10 3 /10 3 CFU/mL, and 10 5 /10 4 CFU/mL against S . agalactiae , S . epidermidis , and S . aureus , respectively. For P . aeruginosa and E . coli , the analytical sensitivity of the multiplex LAMP and qPCR assays were 10 5 /10 4 and 10 6 /10 4 CFU/mL, respectively. The multiplex LAMP assay detects target bacteria without cross-reacting with other bacteria, and exhibited 100% sensitivity and specificity in clinical performance evaluation. Conclusions This ... |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| DOI: | 10.1371/journal.pone.0302783 |
| الاتاحة: | http://dx.doi.org/10.1371/journal.pone.0302783 https://dx.plos.org/10.1371/journal.pone.0302783 |
| Rights: | http://creativecommons.org/licenses/by/4.0/ |
| رقم الانضمام: | edsbas.3A1A6036 |
| قاعدة البيانات: | BASE |
| DOI: | 10.1371/journal.pone.0302783 |
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