GBV-RNA detection by polymerase chain reaction with several primer pairs. 1998

E Quirós, and G Piédrola, and C Maroto
Department of Microbiology, Faculty of Medicine, University of Granada, Spain.

The identification of specific genomic sequences of GB viruses (GBV) has made it possible to utilize the polymerase chain reaction (PCR) for evaluation of the viraemia. Several studies have demonstrated the RNA-GBV presence in sera from different patients amplifying several portions of the virus. In this investigation the PCR results when different regions of GBV (NS3, UTR and putative CORE and E1) were amplified in the same sample. In 245 samples studied there were two (0.8%) discordant results and the NS3 primer showed the greatest sensitivity. The lowest percentage of positivity was obtained with CORE-E1 primers. These results could be because the nucleocapside/E1 region was extremely variable in length and sequences, although degenerated primers and probes were used. Discordances were attributable to laboratory errors, variability in the viral genome, the presence of primer inhibitors in samples or a low viral load.

UI MeSH Term Description Entries
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D012367 RNA, Viral Ribonucleic acid that makes up the genetic material of viruses. Viral RNA
D016133 Polymerase Chain Reaction In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships. Anchored PCR,Inverse PCR,Nested PCR,PCR,Anchored Polymerase Chain Reaction,Inverse Polymerase Chain Reaction,Nested Polymerase Chain Reaction,PCR, Anchored,PCR, Inverse,PCR, Nested,Polymerase Chain Reactions,Reaction, Polymerase Chain,Reactions, Polymerase Chain
D017931 DNA Primers Short sequences (generally about 10 base pairs) of DNA that are complementary to sequences of messenger RNA and allow reverse transcriptases to start copying the adjacent sequences of mRNA. Primers are used extensively in genetic and molecular biology techniques. DNA Primer,Oligodeoxyribonucleotide Primer,Oligodeoxyribonucleotide Primers,Oligonucleotide Primer,Oligonucleotide Primers,Primer, DNA,Primer, Oligodeoxyribonucleotide,Primer, Oligonucleotide,Primers, DNA,Primers, Oligodeoxyribonucleotide,Primers, Oligonucleotide
D018067 Flaviviridae A family of RNA viruses, many of which cause disease in humans and domestic animals. There are three genera FLAVIVIRUS; PESTIVIRUS; and HEPACIVIRUS, as well as several unassigned species.

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