fM to aM nucleic acid amplification for molecular diagnostics in a non-stick-coated metal microfluidic bioreactor.
Sci Rep
; 4: 7344, 2014 Dec 05.
Article
in En
| MEDLINE
| ID: mdl-25475544
ABSTRACT
A sensitive DNA isothermal amplification method for the detection of DNA at fM to aM concentrations for pathogen identification was developed using a non-stick-coated metal microfluidic bioreactor. A portable confocal optical detector was utilized to monitor the DNA amplification in micro- to nanoliter reaction assays in real-time, with fluorescence collection near the optical diffraction limit. The non-stick-coated metal microfluidic bioreactor, with a surface contact angle of 103°, was largely inert to bio-molecules, and DNA amplification could be performed in a minimum reaction volume of 40 nL. The isothermal nucleic acid amplification for Mycoplasma pneumoniae identification in the non-stick-coated microfluidic bioreactor could be performed at a minimum DNA template concentration of 1.3 aM, and a detection limit of three copies of genomic DNA was obtained. This microfluidic bioreactor offers a promising clinically relevant pathogen molecular diagnostic method via the amplification of targets from only a few copies of genomic DNA from a single bacterium.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
DNA, Bacterial
/
Sequence Analysis, DNA
/
Bioreactors
/
Nucleic Acid Amplification Techniques
/
Molecular Diagnostic Techniques
/
Microfluidic Analytical Techniques
Type of study:
Diagnostic_studies
/
Prognostic_studies
Language:
En
Journal:
Sci Rep
Year:
2014
Document type:
Article
Affiliation country:
China