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Comparative Evaluation of LAMP and Nested PCR for the Rapid Diagnosis of Mycobacterium marinum Infection
Infection and Drug Resistance ( IF 2.9 ) Pub Date : 2023-03-20 , DOI: 10.2147/idr.s404929 Yumiao Feng 1, 2 , Miaomiao Wang 2, 3 , Haiqin Jiang 2 , Ying Shi 2 , Wenyue Zhang 2 , Hongsheng Wang 2
Infection and Drug Resistance ( IF 2.9 ) Pub Date : 2023-03-20 , DOI: 10.2147/idr.s404929 Yumiao Feng 1, 2 , Miaomiao Wang 2, 3 , Haiqin Jiang 2 , Ying Shi 2 , Wenyue Zhang 2 , Hongsheng Wang 2
Affiliation
Purpose: Culture of Mycobacterium marinum is very time-consuming, taking several weeks to produce positive results. Seeking rapid and sensitive diagnostic methods for diagnosis can greatly improve patient treatment. Our study aimed to compare the rapid diagnostic abilities of polymerase chain reaction (PCR), nested PCR and loop mediated isothermal amplification (LAMP) of detecting M. marinum in skin samples from patients with M. marinum infection.
Methods: A total of 6 M. marinum strains and 6 skin samples with definite diagnosis of M. marinum infection were included in the study. We optimized LAMP performance for detection of M. marinum genomic DNA and confirmed the specificity of the primers. Then, the sensitivity of the LAMP and nested PCR assays were assessed by M. marinum strains and clinical samples.
Results: Nested PCR was 10-fold more sensitive than the LAMP assay by serial dilution of M. marinum DNA. PCR positive samples were all positive by LAMP detection of 6 clinical M. marinum strains. Out of 6 clinical skin specimens confirmed as M. marinum infection, 0 (0%), 3 (50%), 3 (50%), and 4 (66.6%) were positive by PCR, nested PCR, LAMP and culture. The LAMP shared the same sensitivity than nested PCR in M. marinum strains and clinical samples, but it was easy to perform and faster than nested PCR assay.
Conclusion: Compared with conventional PCR, LAMP and nested PCR are more sensitive and have a higher detection rate of M. marinum in clinical skin specimens. The LAMP assay proved to be more suitable for rapid diagnosis of M. marinum infection in a shorter time, especially in resource-limited settings.
Keywords: Mycobacterium marinum, diagnosis, PCR, nested PCR, LAMP
中文翻译：
LAMP 和巢式 PCR 快速诊断海洋分枝杆菌感染的比较评价
目的：海分枝杆菌的培养非常耗时，需要数周才能产生阳性结果。寻求快速、灵敏的诊断方法进行诊断可以大大提高患者的治疗效果。我们的研究旨在比较聚合酶链反应 (PCR)、巢式 PCR 和环介导等温扩增 (LAMP) 检测海分枝杆菌感染患者皮肤样本中海分枝杆菌的快速诊断能力。方法：共纳入 6 株海分枝杆菌菌株和 6 份明确诊断为海分枝杆菌感染的皮肤样本。我们针对M. marinum的检测优化了 LAMP 性能
基因组 DNA 并验证引物的特异性。然后，通过海分枝杆菌菌株和临床样本评估 LAMP 和巢式 PCR 检测的灵敏度。
结果：巢式 PCR 比通过连续稀释海分枝杆菌DNA的 LAMP 测定法灵敏 10 倍。PCR阳性样本经LAMP检测6株临床海分枝杆菌均呈阳性。在确认为海分枝杆菌感染的 6 份临床皮肤标本中，0 (0%)、3 (50%)、3 (50%) 和 4 (66.6%) 通过 PCR、巢式 PCR、LAMP 和培养呈阳性。LAMP 在海分枝杆菌菌株和临床样本中与巢式 PCR 具有相同的灵敏度，但它比巢式 PCR 测定更容易执行且速度更快。
结论：与常规PCR相比，LAMP和巢式PCR对临床皮肤标本中海分枝杆菌的检出率更高，灵敏度更高。事实证明，LAMP 检测更适合在较短时间内快速诊断海分枝杆菌感染，尤其是在资源有限的环境中。
关键词： 海分枝杆菌，诊断，PCR，巢式 PCR，LAMP
更新日期：2023-03-20
Methods: A total of 6 M. marinum strains and 6 skin samples with definite diagnosis of M. marinum infection were included in the study. We optimized LAMP performance for detection of M. marinum genomic DNA and confirmed the specificity of the primers. Then, the sensitivity of the LAMP and nested PCR assays were assessed by M. marinum strains and clinical samples.
Results: Nested PCR was 10-fold more sensitive than the LAMP assay by serial dilution of M. marinum DNA. PCR positive samples were all positive by LAMP detection of 6 clinical M. marinum strains. Out of 6 clinical skin specimens confirmed as M. marinum infection, 0 (0%), 3 (50%), 3 (50%), and 4 (66.6%) were positive by PCR, nested PCR, LAMP and culture. The LAMP shared the same sensitivity than nested PCR in M. marinum strains and clinical samples, but it was easy to perform and faster than nested PCR assay.
Conclusion: Compared with conventional PCR, LAMP and nested PCR are more sensitive and have a higher detection rate of M. marinum in clinical skin specimens. The LAMP assay proved to be more suitable for rapid diagnosis of M. marinum infection in a shorter time, especially in resource-limited settings.
Keywords: Mycobacterium marinum, diagnosis, PCR, nested PCR, LAMP
中文翻译：
LAMP 和巢式 PCR 快速诊断海洋分枝杆菌感染的比较评价
目的：海分枝杆菌的培养非常耗时，需要数周才能产生阳性结果。寻求快速、灵敏的诊断方法进行诊断可以大大提高患者的治疗效果。我们的研究旨在比较聚合酶链反应 (PCR)、巢式 PCR 和环介导等温扩增 (LAMP) 检测海分枝杆菌感染患者皮肤样本中海分枝杆菌的快速诊断能力。方法：共纳入 6 株海分枝杆菌菌株和 6 份明确诊断为海分枝杆菌感染的皮肤样本。我们针对M. marinum的检测优化了 LAMP 性能
基因组 DNA 并验证引物的特异性。然后，通过海分枝杆菌菌株和临床样本评估 LAMP 和巢式 PCR 检测的灵敏度。
结果：巢式 PCR 比通过连续稀释海分枝杆菌DNA的 LAMP 测定法灵敏 10 倍。PCR阳性样本经LAMP检测6株临床海分枝杆菌均呈阳性。在确认为海分枝杆菌感染的 6 份临床皮肤标本中，0 (0%)、3 (50%)、3 (50%) 和 4 (66.6%) 通过 PCR、巢式 PCR、LAMP 和培养呈阳性。LAMP 在海分枝杆菌菌株和临床样本中与巢式 PCR 具有相同的灵敏度，但它比巢式 PCR 测定更容易执行且速度更快。
结论：与常规PCR相比，LAMP和巢式PCR对临床皮肤标本中海分枝杆菌的检出率更高，灵敏度更高。事实证明，LAMP 检测更适合在较短时间内快速诊断海分枝杆菌感染，尤其是在资源有限的环境中。
关键词： 海分枝杆菌，诊断，PCR，巢式 PCR，LAMP
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