Development of a magnetic bead microarray for simultaneous and simple detection of four pestiviruses.
This study reports a novel method for the rapid detection and identification of the four recognized species in the pestivirus genus of the Flaviviridae family, i.e. classical swine fever virus (CSFV), border disease virus (BDV), bovine viral diarrhoea virus type 1 (BVDV1) and type 2 (BVDV2). The analysis of pestivirus PCR products was performed on microarrays by means of magnetic bead detection. The process utilizes an oligonucleotide array, onto which 5' biotinylated PCR products were hybridized, followed by visualization with streptavidin-coated magnetic particles by the naked eye, microscope or biochip reader. The assay was tested on a collection of pestiviruses that included all four species and allowed a specific and sensitive detection. Sensitivity was compared with other post-PCR detection methods, namely gel electrophoresis and suspension microarray. The results indicate that due to its high sensitivity, specificity and simple detection procedure, the magnetic bead assay provides a powerful tool for detection and identification of viral pathogens. Considering the simplicity of the assay, the protocols for hybridization and magnetic bead detection offer an emerging application for molecular diagnoses in virology that is amenable for use in a modestly equipped laboratory.
Joint R&D Division, Departments of Virology, The National Veterinary Institute (SVA) and The Swedish University of Agricultural Sciences (SLU), SE-751 89, Uppsala, Sweden.
Neil
N
Leblanc
Jesper
J
Gantelius
Jochen M
JM
Schwenk
Karl
K
Ståhl
Jonas
J
Blomberg
Helene
H
Andersson-Svahn
Sándor
S
Belák
D000818:Animals; D018083:Border disease virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D002417:Cattle; D006692:Classical Swine Fever Virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D030241:Diarrhea Virus 1, Bovine Viral / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D030242:Diarrhea Virus 2, Bovine Viral / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D008280:Magnetics; D008863:Microspheres; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010576:Pestivirus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013552:Swine