JEV-nanobarcode and colorimetric reverse transcription loop-mediated isothermal amplification (cRT-LAMP)
| Autor: | Ji-Young Ahn, Min-Suk Song, Gna Ahn, Yang-Hoon Kim, Se Hee Lee, Beom-Ku Han |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Swine
viruses Immobilized Nucleic Acids Loop-mediated isothermal amplification Metal Nanoparticles Urine Dengue virus medicine.disease_cause Virus Analytical Chemistry Limit of Detection medicine Animals Humans Nucleic Acid Amplification Tests Reverse Transcription Loop-mediated Isothermal Amplification Encephalitis Virus Japanese Detection limit Base Sequence Chemistry Japanese encephalitis Amplicon medicine.disease Virology Blood Molecular Diagnostic Techniques RNA Viral Colorimetry Gold Poly A Nucleic Acid Amplification Techniques |
| Zdroj: | Microchimica Acta. 188 |
| ISSN: | 1436-5073 0026-3672 |
| DOI: | 10.1007/s00604-021-04986-9 |
| Popis: | Nucleic acid amplification tests (NAATs) are powerful tools for the Japanese encephalitis virus (JEV). We demonstrated highly sensitive, specific, and rapid detection of JEV by colorimetric reverse-transcription loop-mediated isothermal amplification (cRT-LAMP). Under optimized conditions, the RT-LAMP assay results showed that the limit of detection was approximately equivalent to 1 RNA genome copy/μL with an assay time of 30 min. The assay was highly specific to JEV when tested with other mosquito-borne virus panels (Zika virus and dengue virus types 2–4). The ability to detect JEV directly from crude human sample matrices (serum and urine) demonstrated the suitability of our JEV RT-LAMP for widespread clinical application. The JEV RT-LAMP provides combination of rapid colorimetric determination of true-positive JEV RT-LAMP amplicons with our recently developed JEV-nanobarcodes, measured at absorbance wavelenght of 530 (A530) and 650 (A650), which have a limit of detection of 23.3 ng/μL. The AuNP:polyA10-JEV RT-LAMP nanobarcodes exhibited superior capability for stabilizing the true-positive JEV RT-LAMP amplicons against salt-induced AuNP aggregation, which improved the evaluation of true/false positive signals in the assay. These advances enable to expand the use of RT-LAMP for point-of-care tests, which will greatly bolster JEV clinical programs. |
| Databáze: | OpenAIRE |
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