A Simple and Multiplex Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid Detection of SARS-CoV
| Autor: | Minhee Kang, Jiyeon Kim, Eunkyoung Park, Eung Soo Hwang, Jin Hwa Kim, Doo Ryeon Chung |
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| Rok vydání: | 2019 |
| Předmět: |
Biomedical Engineering
Loop-mediated isothermal amplification Bioengineering 02 engineering and technology Biology medicine.disease_cause 01 natural sciences Rapid detection medicine Multiplex Electrical and Electronic Engineering Point-of-care test Gene Coronavirus Detection limit 010401 analytical chemistry SARS-CoV 021001 nanoscience & nanotechnology Virology 0104 chemical sciences Standard curve Open reading frame Original Article 0210 nano-technology Colorimetric detection Biotechnology |
| Zdroj: | Biochip Journal |
| ISSN: | 2092-7843 1976-0280 |
| Popis: | The current diagnosis of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) relies on laboratory-based tests since its clinical features are nonspecific, unlike other respiratory pathogens. Therefore, the development of a rapid and simple method for on-site detection of SARS-CoV is crucial for the identification and prevention of future SARS outbreaks. In this study, a simple colorimetric and multiplex loop-mediated isothermal amplification (LAMP) assay was developed to rapid screening of severe acute respiratory syndrome-associated coronavirus (SARS-CoV). It can be visually detected based on color change and monitored in real-time with fluorescent signals. The performance of this assay, based on six primers targeting open reading frame (ORF1b) and nucleocapsid (N) genes located in different regions of the SARS-CoV, was compared with real-time RT-PCR assay using various concentrations of target genes. The detection limit of the LAMP assay was comparable to that of real-time RT-PCR assay and therefore a few target RNA to 104-105 copies could be detected within a short period of time (20–25 min). In addition, we established a multiplex real-time LAMP assay to simultaneously detect two target regions within the SARS-CoV genome. Two target sequences were amplified by specific primers in the same reaction tube and revealed that it was able to detect down to 105 copies. The standard curve had a linear relationship with similar amplification efficiencies. The LAMP assay results in shorter “sample-to-answer” time than conventional PCR method. Therefore, it is suitable not only for diagnosis of clinical test, but also for surveillance of SARS virus in developing countries. Electronic Supplementary Material Supplementary material is available for this article at 10.1007/s13206-019-3404-3 and is accessible for authorized users. |
| Databáze: | OpenAIRE |
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