Method for colorimetric detection of double-stranded nucleic acid using leuco triphenylmethane dyes.
Anal Biochem
; 473: 28-33, 2015 Mar 15.
Article
em En
| MEDLINE
| ID: mdl-25575759
ABSTRACT
Because loop-mediated isothermal amplification (LAMP) can amplify substantial amounts of DNA under isothermal conditions, its applications for simple genetic testing have attracted considerable attention. A positive LAMP reaction is indicated by the turbidity caused by by-products or by the color change after adding a metallochromic indicator to the reaction solution, but these methods have certain limitations. Leuco crystal violet (LCV), a colorless dye obtained after sodium sulfite treatment of crystal violet (CV), was used as a new colorimetric method for detecting LAMP. LCV is reconverted into CV through contact with double-stranded DNA (dsDNA). Therefore, the positive reaction of LAMP is indicated by color change from colorless to violet. The assay is sensitive enough to detect LAMP products, with a detection limit of 7.1 ng/µl for dsDNA. It is also highly selective to dsDNA, and interference with single-stranded DNA and deoxynucleotide triphosphates (dNTPs) is not observed. LCV facilitates direct colorimetric detection of the main product rather than a by-product of the LAMP reaction; therefore, this method can be used under various reaction conditions such as those with added pyrophosphatase in solution. This colorimetric LAMP detection method using LCV is useful for point-of-care genetic testing given its simplicity.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Compostos de Tritil
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DNA
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Colorimetria
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Corantes
Tipo de estudo:
Diagnostic_studies
Idioma:
En
Ano de publicação:
2015
Tipo de documento:
Article