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JEV-nanobarcode and colorimetric reverse transcription loop-mediated isothermal amplification (cRT-LAMP).
- Source :
-
Mikrochimica acta [Mikrochim Acta] 2021 Sep 08; Vol. 188 (10), pp. 333. Date of Electronic Publication: 2021 Sep 08. - Publication Year :
- 2021
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Abstract
- 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 (A <subscript>530</subscript> ) and 650 (A <subscript>650</subscript> ), which have a limit of detection of 23.3 ng/μL. The AuNP:polyA <subscript>10</subscript> -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. The JEV RT-LAMP nanobarcode assay targeting the envelope (E) gene and MgSO <subscript>4</subscript> induced AuNP aggregation, indicated by an instant pink-to-violet colorimetric read-out.<br /> (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)
- Subjects :
- Animals
Base Sequence
Blood virology
Gold chemistry
Humans
Immobilized Nucleic Acids chemistry
Limit of Detection
Metal Nanoparticles chemistry
Poly A chemistry
RNA, Viral blood
RNA, Viral urine
Swine
Urine virology
Colorimetry methods
Encephalitis Virus, Japanese chemistry
Molecular Diagnostic Techniques methods
Nucleic Acid Amplification Techniques methods
RNA, Viral analysis
Subjects
Details
- Language :
- English
- ISSN :
- 1436-5073
- Volume :
- 188
- Issue :
- 10
- Database :
- MEDLINE
- Journal :
- Mikrochimica acta
- Publication Type :
- Academic Journal
- Accession number :
- 34498149
- Full Text :
- https://doi.org/10.1007/s00604-021-04986-9