Evaluation of a Commercial Multiplex Real-Time PCR with Melting Curve Analysis for the Detection of <i>Mycobacterium tuberculosis</i> Complex and Five Nontuberculous Mycobacterial Species
Background: Accurate and timely diagnosis of mycobacterial infections, including <i>Mycobacterium tuberculosis</i> complex (MTBC) and nontuberculous mycobacteria (NTM), is crucial for effective disease management. Methods: This study evaluated the performance of the NeoPlex TB/NTM-5 Dete...
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2024-12-01
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| author | Keun Ju Kim Yunhee Chang Seung Gyu Yun Myung-Hyun Nam Yunjung Cho |
| author_facet | Keun Ju Kim Yunhee Chang Seung Gyu Yun Myung-Hyun Nam Yunjung Cho |
| author_sort | Keun Ju Kim |
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| description | Background: Accurate and timely diagnosis of mycobacterial infections, including <i>Mycobacterium tuberculosis</i> complex (MTBC) and nontuberculous mycobacteria (NTM), is crucial for effective disease management. Methods: This study evaluated the performance of the NeoPlex TB/NTM-5 Detection Kit (NeoPlex assay, Seongnam, Republic of Korea), a multiplex real-time PCR assay that incorporates melting curve analysis, compared with the line-probe assay (LPA). The NeoPlex assay could simultaneously detect and differentiate MTBC from five other NTM species: <i>Mycobacterium intracellulare</i>, <i>Mycobacterium avium</i>, <i>Mycobacterium kansasii</i>, <i>Mycobacterium abscessus</i>, and <i>Mycobacterium massiliense</i>. A total of 91 acid-fast bacillus culture-positive samples, comprising 36 MTBC and 55 NTM isolates, were collected from the Korea University Anam Hospital. Results: The NeoPlex assay successfully detected nucleic acids in 87 of the 91 isolates (95.6%). Notably, it identified additional mycobacterial nucleic acids not detected by the LPA in eight isolates. These findings were confirmed via DNA sequencing. The assay had 100% sensitivity and specificity for <i>M. intracellulare</i>, <i>M. abscessus</i>, <i>M. massilense</i>, NTM, and MTBC, whereas it had 100% specificity and sensitivity of 90.9% and 75.0% for <i>M. avium</i> and <i>M. kansasii</i>, respectively. Conclusions: These results highlight the potential of the NeoPlex assay to enhance rapid and accurate diagnosis of mycobacterial infections, particularly in settings in which prompt treatment initiation is essential. |
| format | Article |
| id | doaj-art-3001ce6fb37e45aeb462456b16c67352 |
| institution | Kabale University |
| issn | 2076-2607 |
| language | English |
| publishDate | 2024-12-01 |
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| series | Microorganisms |
| spelling | doaj-art-3001ce6fb37e45aeb462456b16c673522025-01-24T13:42:20ZengMDPI AGMicroorganisms2076-26072024-12-011312610.3390/microorganisms13010026Evaluation of a Commercial Multiplex Real-Time PCR with Melting Curve Analysis for the Detection of <i>Mycobacterium tuberculosis</i> Complex and Five Nontuberculous Mycobacterial SpeciesKeun Ju Kim0Yunhee Chang1Seung Gyu Yun2Myung-Hyun Nam3Yunjung Cho4Department of Laboratory Medicine, Korea University Anam Hospital, Korea University College of Medicine, Seoul 02841, Republic of KoreaDepartment of Biomedical Laboratory Science, Kyungnam College of Information & Technology, Busan 47011, Republic of KoreaDepartment of Laboratory Medicine, Korea University Anam Hospital, Korea University College of Medicine, Seoul 02841, Republic of KoreaDepartment of Laboratory Medicine, Korea University Anam Hospital, Korea University College of Medicine, Seoul 02841, Republic of KoreaDepartment of Laboratory Medicine, Korea University Anam Hospital, Korea University College of Medicine, Seoul 02841, Republic of KoreaBackground: Accurate and timely diagnosis of mycobacterial infections, including <i>Mycobacterium tuberculosis</i> complex (MTBC) and nontuberculous mycobacteria (NTM), is crucial for effective disease management. Methods: This study evaluated the performance of the NeoPlex TB/NTM-5 Detection Kit (NeoPlex assay, Seongnam, Republic of Korea), a multiplex real-time PCR assay that incorporates melting curve analysis, compared with the line-probe assay (LPA). The NeoPlex assay could simultaneously detect and differentiate MTBC from five other NTM species: <i>Mycobacterium intracellulare</i>, <i>Mycobacterium avium</i>, <i>Mycobacterium kansasii</i>, <i>Mycobacterium abscessus</i>, and <i>Mycobacterium massiliense</i>. A total of 91 acid-fast bacillus culture-positive samples, comprising 36 MTBC and 55 NTM isolates, were collected from the Korea University Anam Hospital. Results: The NeoPlex assay successfully detected nucleic acids in 87 of the 91 isolates (95.6%). Notably, it identified additional mycobacterial nucleic acids not detected by the LPA in eight isolates. These findings were confirmed via DNA sequencing. The assay had 100% sensitivity and specificity for <i>M. intracellulare</i>, <i>M. abscessus</i>, <i>M. massilense</i>, NTM, and MTBC, whereas it had 100% specificity and sensitivity of 90.9% and 75.0% for <i>M. avium</i> and <i>M. kansasii</i>, respectively. Conclusions: These results highlight the potential of the NeoPlex assay to enhance rapid and accurate diagnosis of mycobacterial infections, particularly in settings in which prompt treatment initiation is essential.https://www.mdpi.com/2076-2607/13/1/26tuberculosismelting curve analysisNTM differentiationNeoPlex TB/NTM |
| spellingShingle | Keun Ju Kim Yunhee Chang Seung Gyu Yun Myung-Hyun Nam Yunjung Cho Evaluation of a Commercial Multiplex Real-Time PCR with Melting Curve Analysis for the Detection of <i>Mycobacterium tuberculosis</i> Complex and Five Nontuberculous Mycobacterial Species Microorganisms tuberculosis melting curve analysis NTM differentiation NeoPlex TB/NTM |
| title | Evaluation of a Commercial Multiplex Real-Time PCR with Melting Curve Analysis for the Detection of <i>Mycobacterium tuberculosis</i> Complex and Five Nontuberculous Mycobacterial Species |
| title_full | Evaluation of a Commercial Multiplex Real-Time PCR with Melting Curve Analysis for the Detection of <i>Mycobacterium tuberculosis</i> Complex and Five Nontuberculous Mycobacterial Species |
| title_fullStr | Evaluation of a Commercial Multiplex Real-Time PCR with Melting Curve Analysis for the Detection of <i>Mycobacterium tuberculosis</i> Complex and Five Nontuberculous Mycobacterial Species |
| title_full_unstemmed | Evaluation of a Commercial Multiplex Real-Time PCR with Melting Curve Analysis for the Detection of <i>Mycobacterium tuberculosis</i> Complex and Five Nontuberculous Mycobacterial Species |
| title_short | Evaluation of a Commercial Multiplex Real-Time PCR with Melting Curve Analysis for the Detection of <i>Mycobacterium tuberculosis</i> Complex and Five Nontuberculous Mycobacterial Species |
| title_sort | evaluation of a commercial multiplex real time pcr with melting curve analysis for the detection of i mycobacterium tuberculosis i complex and five nontuberculous mycobacterial species |
| topic | tuberculosis melting curve analysis NTM differentiation NeoPlex TB/NTM |
| url | https://www.mdpi.com/2076-2607/13/1/26 |
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