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Synthetic oligonucleotide cocktails as probes for detection of human parvovirus B19. 1995

H A Cubie, and J Grzybowski, and C da Silva, and L Duncan, and T Brown, and N M Smith
Regional Virus Laboratory, City Hospital, Edinburgh, Scotland.

A cocktail of 10 oligonucleotides selected at intervals along the length of the genome of human parvovirus B19 was labelled enzymically with digoxigenin and chemically with either digoxigenin (DIG) or dinitrophenyl (DNP). Chemical labelling was easier and more practical for the production of large quantities of probe. Pools labelled with either digoxigenin or DNP could detect 10 fg of B19 DNA in a dot blot reaction using an alkaline phosphatase antibody conjugate and colorimetric detection. Formalin fixed tissue from 11 consecutive cases of fetal hydrops were examined by in situ hybridisation (ISH). Both probe cocktails detected human parvovirus B19 DNA in 3 cases, with positive cells in all tissues examined and with equal sensitivity. The DNP pool is significantly cheaper and simpler to produce and could provide an inexpensive reagent suitable for diagnostic detection of viral nucleic acid in histopathological material.

UI MeSH Term Description Entries
D008969 Molecular Sequence Data Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories. Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D004076 Digoxigenin 3 beta,12 beta,14-Trihydroxy-5 beta-card-20(22)-enolide. A cardenolide which is the aglycon of digoxin. Can be obtained by hydrolysis of digoxin or from Digitalis orientalis L. and Digitalis lanata Ehrh. Lanadigenin
D004136 Dinitrobenzenes Benzene derivatives which are substituted with two nitro groups in the ortho, meta or para positions. Dinitrobenzene,Dinitrophenyl Compound,Dinitrophenyl Compounds,Dinitrotoluene,Dinitrotoluenes,Compound, Dinitrophenyl
D004279 DNA, Viral Deoxyribonucleic acid that makes up the genetic material of viruses. Viral DNA
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D001483 Base Sequence The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence. DNA Sequence,Nucleotide Sequence,RNA Sequence,DNA Sequences,Base Sequences,Nucleotide Sequences,RNA Sequences,Sequence, Base,Sequence, DNA,Sequence, Nucleotide,Sequence, RNA,Sequences, Base,Sequences, DNA,Sequences, Nucleotide,Sequences, RNA
D012680 Sensitivity and Specificity Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed) Specificity,Sensitivity,Specificity and Sensitivity
D015160 Hydrops Fetalis Abnormal accumulation of serous fluid in two or more fetal compartments, such as SKIN; PLEURA; PERICARDIUM; PLACENTA; PERITONEUM; AMNIOTIC FLUID. General fetal EDEMA may be of non-immunologic origin, or of immunologic origin as in the case of ERYTHROBLASTOSIS FETALIS. Edema, Fetal,Fetal Edema,Fetal Hydrops,Familial Non-Immune Hydrops Fetalis,Hydrops Fetalis Nonimmune,Hydrops Fetalis, Idiopathic,Hydrops Fetalis, Immune,Hydrops Fetalis, Non-Immune,Hydrops Fetalis, Nonimmune,Idiopathic Hydrops Fetalis,Immune Hydrops Fetalis,Non-Immune Hydrops Fetalis,Familial Non Immune Hydrops Fetalis,Fetali, Idiopathic Hydrops,Fetalis Nonimmune, Hydrops,Fetalis Nonimmunes, Hydrops,Fetalis, Idiopathic Hydrops,Hydrops Fetali, Idiopathic,Hydrops Fetali, Nonimmune,Hydrops Fetalis Nonimmunes,Hydrops, Fetal,Idiopathic Hydrops Fetali,Nonimmune Hydrops Fetali,Nonimmune Hydrops Fetalis,Nonimmune, Hydrops Fetalis,Nonimmunes, Hydrops Fetalis
D015345 Oligonucleotide Probes Synthetic or natural oligonucleotides used in hybridization studies in order to identify and study specific nucleic acid fragments, e.g., DNA segments near or within a specific gene locus or gene. The probe hybridizes with a specific mRNA, if present. Conventional techniques used for testing for the hybridization product include dot blot assays, Southern blot assays, and DNA:RNA hybrid-specific antibody tests. Conventional labels for the probe include the radioisotope labels 32P and 125I and the chemical label biotin. Oligodeoxyribonucleotide Probes,Oligonucleotide Probe,Oligoribonucleotide Probes,Probe, Oligonucleotide,Probes, Oligodeoxyribonucleotide,Probes, Oligonucleotide,Probes, Oligoribonucleotide
D016732 Parvovirus B19, Human The type species of ERYTHROVIRUS and the etiological agent of ERYTHEMA INFECTIOSUM, a disease most commonly seen in school-age children. B19 virus,Human Parvovirus B19,Parvovirus B19,B19 viruses,B19, Parvovirus,Parvovirus B19s

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