Biotin-psolaren labeled cDNA amplicons for genotyping rotavirus strains by dot hybridization assay.
A simple method to produce biotinylated probes by utilizing an amine-psoralen to conjugate an ester-biotin to nucleic acid molecules is described. It is simple, rapid, and well suited to label cDNA probes with any PCR-generated amplicon for hybridization assay. Its application to identify or to confirm the G and P genotypes of rotavirus-derived amplicons is described; however, it may be used to label amplicons of any origin. As an alternative or as a complementary test to either PCR-typing assay and/or sequencing, it should reduce considerably the laboratory costs required to genotype fully virus strains in large epidemiological surveys conducted in developing countries.
Domingues
André Luiz S
AL
Departmento de Virologia, Instituto de Microbiologia, Universidade Federal do Rio de Janeiro, Cidade Universitária, CCS, Bloco I, Ilha do Fundão, Rio de Janeiro, RJ 21941-590, Brazil. andredomingues@dcb.uftm.edu.br
Silva
Maria Helena
MH
Gouvea
Vera
V
D001710:Biotin / Q000378:metabolism*; D019921:Biotinylation; D015342:DNA Probes; D018076:DNA, Complementary / Q000235:genetics; D005363:Ficusin / Q000378:metabolism*; D016679:Genome, Viral; D005838:Genotype; D015336:Molecular Probe Techniques; D009693:Nucleic Acid Hybridization; D012401:Rotavirus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*
D015342:DNA Probes; D018076:DNA, Complementary; D001710:Biotin; D005363:Ficusin