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Rolling circle amplification based colorimetric determination of Staphylococcus aureus.
- Source :
- Microchimica Acta; Feb2020, Vol. 187 Issue 2, p1-10, 10p
- Publication Year :
- 2020
-
Abstract
- A colorimetric microplate assay for determination of Staphylococcus aureus DNA is described. Linear padlock probes were designed to recognize target sequences. After DNA binding, the linear padlock probes were circularized by ligation and then hybridize with biotin-labeled capture probes. Biotin-labeled capture probes act as primers to initiate the RCA. The biotin-labeled RCA products hybridize with digoxin-labeled signal probes fixed on streptavidin-functionalized wells of a 96-well plate. To enhance sensitivity, an AuNP-anti-digoxigenin-POx-HRP conjugate was added to the wells and then bound to digoxin-labeled signalling probes. The oxidation of tetramethylbenzidine (TMB) by H<subscript>2</subscript>O<subscript>2</subscript> produces a color change from colorless to blue via HRP catalysis. After the reaction was terminated, absorbance is measured at 450 nm. For target sequences of Staphylococcus aureus, the detection limit is 1.2 pM. For genomic DNA, the detection limit is 7.4 pg.μL<superscript>−1</superscript>. The potential application of the method was verified by analyzing spiked food samples. [ABSTRACT FROM AUTHOR]
- Subjects :
- STAPHYLOCOCCUS aureus
DETECTION limit
DNA primers
CIRCLE
DNA
GOLD nanoparticles
Subjects
Details
- Language :
- English
- ISSN :
- 00263672
- Volume :
- 187
- Issue :
- 2
- Database :
- Complementary Index
- Journal :
- Microchimica Acta
- Publication Type :
- Academic Journal
- Accession number :
- 141879764
- Full Text :
- https://doi.org/10.1007/s00604-019-4082-5