Simple Point-of-Care Nucleic Acid Amplification Test for Rapid SARS-CoV-2 Infection Diagnosis
After three years of the SARS-CoV-2 pandemic, the demand for developing field-deployable point-of-care (PoC) molecular diagnostic tests has increased. Although RT-qPCR is the molecular diagnostic gold standard and is accurate, it is not readily applied to point-of-care testing (POCT). Meanwhile, rap...
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MDPI AG
2023-09-01
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| Series: | Diagnostics |
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| Online Access: | https://www.mdpi.com/2075-4418/13/18/3001 |
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| author | Hyunseul Jee Minkyeong Choi In Su Park Junmin Lee Woong Sik Jang Chae Seung Lim |
| author_facet | Hyunseul Jee Minkyeong Choi In Su Park Junmin Lee Woong Sik Jang Chae Seung Lim |
| author_sort | Hyunseul Jee |
| collection | DOAJ |
| description | After three years of the SARS-CoV-2 pandemic, the demand for developing field-deployable point-of-care (PoC) molecular diagnostic tests has increased. Although RT-qPCR is the molecular diagnostic gold standard and is accurate, it is not readily applied to point-of-care testing (POCT). Meanwhile, rapid diagnostic kits have the disadvantage of low sensitivity. Recently, rapid isothermal nucleic acid amplification technology has emerged as an alternative for rapid diagnosis. Here, we developed a rapid SARS-CoV-2 reverse transcription loop-mediated isothermal amplification (RT-LAMP)-lateral flow assay (LFA) kit. This kit includes a Chelex-100/boiling nucleic acid extraction device and a one-step amplification detection apparatus capable of performing the entire process, from RNA extraction to detection, and diagnosing SARS-CoV-2 infection within 40 min without contamination. The detection limits of the rapid SARS-CoV-2 RT-LAMP-LFA kit were 100 plaque-forming units (PFUs) mL<sup>−1</sup> and 10<sup>−1</sup> PFU mL<sup>−1</sup> for RNA samples extracted using the Chelex-100/boiling nucleic acid extraction device and commercial AdvansureTM E3 system, respectively. The sensitivity and specificity of the rapid SARS-CoV-2 RT-LAMP-LFA kit were 97.8% and 100%, respectively. Our SARS-CoV-2 RT-LAMP-LFA kit exhibited high sensitivity and specificity within 40 min without requiring laboratory instruments, suggesting that the kit could be used as a rapid POC molecular diagnostic test for SARS-CoV-2. |
| first_indexed | 2024-03-10T22:52:21Z |
| format | Article |
| id | doaj.art-a4d7e5f9878b4f29bc8aaed3122fa5c2 |
| institution | Directory Open Access Journal |
| issn | 2075-4418 |
| language | English |
| last_indexed | 2024-03-10T22:52:21Z |
| publishDate | 2023-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Diagnostics |
| spelling | doaj.art-a4d7e5f9878b4f29bc8aaed3122fa5c22023-11-19T10:14:32ZengMDPI AGDiagnostics2075-44182023-09-011318300110.3390/diagnostics13183001Simple Point-of-Care Nucleic Acid Amplification Test for Rapid SARS-CoV-2 Infection DiagnosisHyunseul Jee0Minkyeong Choi1In Su Park2Junmin Lee3Woong Sik Jang4Chae Seung Lim5BK21 Graduate Program, Department of Biomedical Sciences, College of Medicine, Korea University, Seoul 02841, Republic of KoreaBK21 Graduate Program, Department of Biomedical Sciences, College of Medicine, Korea University, Seoul 02841, Republic of KoreaBK21 Graduate Program, Department of Biomedical Sciences, College of Medicine, Korea University, Seoul 02841, Republic of KoreaBK21 Graduate Program, Department of Biomedical Sciences, College of Medicine, Korea University, Seoul 02841, Republic of KoreaEmergency Medicine, College of Medicine, Korea University Guro Hospital, Seoul 08308, Republic of KoreaDepartment of Laboratory Medicine, College of Medicine, Korea University Guro Hospital, Seoul 08308, Republic of KoreaAfter three years of the SARS-CoV-2 pandemic, the demand for developing field-deployable point-of-care (PoC) molecular diagnostic tests has increased. Although RT-qPCR is the molecular diagnostic gold standard and is accurate, it is not readily applied to point-of-care testing (POCT). Meanwhile, rapid diagnostic kits have the disadvantage of low sensitivity. Recently, rapid isothermal nucleic acid amplification technology has emerged as an alternative for rapid diagnosis. Here, we developed a rapid SARS-CoV-2 reverse transcription loop-mediated isothermal amplification (RT-LAMP)-lateral flow assay (LFA) kit. This kit includes a Chelex-100/boiling nucleic acid extraction device and a one-step amplification detection apparatus capable of performing the entire process, from RNA extraction to detection, and diagnosing SARS-CoV-2 infection within 40 min without contamination. The detection limits of the rapid SARS-CoV-2 RT-LAMP-LFA kit were 100 plaque-forming units (PFUs) mL<sup>−1</sup> and 10<sup>−1</sup> PFU mL<sup>−1</sup> for RNA samples extracted using the Chelex-100/boiling nucleic acid extraction device and commercial AdvansureTM E3 system, respectively. The sensitivity and specificity of the rapid SARS-CoV-2 RT-LAMP-LFA kit were 97.8% and 100%, respectively. Our SARS-CoV-2 RT-LAMP-LFA kit exhibited high sensitivity and specificity within 40 min without requiring laboratory instruments, suggesting that the kit could be used as a rapid POC molecular diagnostic test for SARS-CoV-2.https://www.mdpi.com/2075-4418/13/18/3001point-of-careSARS-CoV-2loop-mediated isothermal amplificationnucleic acid lateral flow |
| spellingShingle | Hyunseul Jee Minkyeong Choi In Su Park Junmin Lee Woong Sik Jang Chae Seung Lim Simple Point-of-Care Nucleic Acid Amplification Test for Rapid SARS-CoV-2 Infection Diagnosis Diagnostics point-of-care SARS-CoV-2 loop-mediated isothermal amplification nucleic acid lateral flow |
| title | Simple Point-of-Care Nucleic Acid Amplification Test for Rapid SARS-CoV-2 Infection Diagnosis |
| title_full | Simple Point-of-Care Nucleic Acid Amplification Test for Rapid SARS-CoV-2 Infection Diagnosis |
| title_fullStr | Simple Point-of-Care Nucleic Acid Amplification Test for Rapid SARS-CoV-2 Infection Diagnosis |
| title_full_unstemmed | Simple Point-of-Care Nucleic Acid Amplification Test for Rapid SARS-CoV-2 Infection Diagnosis |
| title_short | Simple Point-of-Care Nucleic Acid Amplification Test for Rapid SARS-CoV-2 Infection Diagnosis |
| title_sort | simple point of care nucleic acid amplification test for rapid sars cov 2 infection diagnosis |
| topic | point-of-care SARS-CoV-2 loop-mediated isothermal amplification nucleic acid lateral flow |
| url | https://www.mdpi.com/2075-4418/13/18/3001 |
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