Document Detail


Portable chemiluminescence multiplex biosensor for quantitative detection of three B19 DNA genotypes.
MedLine Citation:
PMID:  23187829     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
A miniaturized multiplex biosensor exploiting a microfluidic oligonucleotide array and chemiluminescence (CL) lensless imaging detection has been developed for parvovirus B19 genotyping. The portable device consists of a reaction chip, comprising a glass slide arrayed with three B19 genotype-specific probes and coupled with a polydimethylsiloxane microfluidic layer, and a charge-coupled device camera modified for lensless CL imaging. Immobilized probes were used in DNA hybridization reactions with biotin-labeled targets, and then hybrids were measured by means of an avidin-horseradish peroxidase (HRP) conjugate and CL detection. All hybridization assay procedures have been optimized to be performed at room temperature through the microfluidic elements of the reaction chip, with sample and reagents delivery via capillary force exploiting adsorbent pads to drive fluids along the microchannels. The biosensor enabled multiplex detection of all B19 genotypes, with detectability down to 80 pmol L(-1) for all B19 genotype oligonucleotides and 650 pmol L(-1) for the amplified product of B19 genotype 1, which is comparable with that obtained in traditional PCR-ELISA formats and with notably shorter assay time (30 min vs. 2 h). The specificity of the assay has been evaluated by performing DNA-DNA hybridization reactions among sequences with different degrees of homology, and no cross hybridizations among B19 genotypes have been observed. The clinical applicability has been demonstrated by assaying amplified products obtained from B19 reference serum samples, with results completely consistent with the reference PCR-ELISA method. The next crucial step will be integration in the biosensor of a miniaturized PCR system for DNA amplification and for heat treatment of amplified products.
Authors:
Mara Mirasoli; Francesca Bonvicini; Luisa Stella Dolci; Martina Zangheri; Giorgio Gallinella; Aldo Roda
Related Documents :
1738139 - Structure and dna binding activity of analogues of 1,5-bis(4-amidinophenoxy)pentane (pe...
10197039 - Cytotoxics derived from distamycin a and congeners.
1315539 - Selective binding to at sequences in dna by an acridine-linked peptide containing the s...
9298959 - Structure-based discovery of ligands targeted to the rna double helix.
9397169 - Boron-containing polyamines as dna targeting agents for neutron capture therapy of brai...
17506529 - Unusually strong binding to the dna minor groove by a highly twisted benzimidazole diph...
11832959 - Plasmodium, human and anopheles genomics and malaria.
15583029 - Dna replication checkpoint control of wee1 stability by vertebrate hsl7.
23831579 - Nmr structure of the n-terminal-most hrdc1 domain of recq helicase from deinococcus rad...
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-11-28
Journal Detail:
Title:  Analytical and bioanalytical chemistry     Volume:  -     ISSN:  1618-2650     ISO Abbreviation:  Anal Bioanal Chem     Publication Date:  2012 Nov 
Date Detail:
Created Date:  2012-11-28     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101134327     Medline TA:  Anal Bioanal Chem     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
Department of Chemistry "G. Ciamician", University of Bologna, Via Selmi 2, 40126, Bologna, Italy, mara.mirasoli@unibo.it.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Time-course measurements of caffeine and its metabolites extracted from fingertips after coffee inta...
Next Document:  Expression and role of GLUT-1, MCT-1, and MCT-4 in malignant pleural mesothelioma.