Detection of B19 parvovirus in human fetal tissues by in situ hybridisation

Abstract

Evidence of B19 parvovirus infection was sought by in situ hybridisation with biotinylated probes in 65 tissue samples from 32 pregnancies (fetuses, products of conception and/or placentas). Twenty-seven samples were reactive and the results were confirmed by other methods for B19 virus detection in 22 cases. The other methods used were in situ hybridisation with 3H and 35S labelled probes; dot-blot hybridisation with biotin and 32P labelled probes; polymerase chain reaction assay; negative stain and thin section electron microscopy; and radioimmunoassay for B19 antigen. The five false positive results by in situ hybridisation with biotinylated probes were considered to be due to non-specific biotin capture and were more frequent with unfixed samples than with formalin fixed material. It was concluded that while biotinylated probes offered advantages over radioactive probes for detecting B19 DNA by in situ hybridisation, positive findings should be confirmed by other methods.

Prenatal diagnosis of congenital parvovirus B19 infection: value of serological and PCR techniques in maternal and fetal serum

“…Maternal infection during pregnancy with human parvovirus B19 (B19) is a serious cause of fetal morbidity and mortality because of the virus' ability to cross the placenta and to replicate lytically in fetal erythroid precursor cells thereby interrupting fetal erythropoiesis (Knott et al, 1984;Ozawa et al, 1986;Anderson et al, 1988;Nascimento et al, 1991). Severe anaemia and hypoxia, followed by vascular dysfunction, can result in secondary heart failure leading to fetal death or non-immune hydrops fetalis (NIHF) (Brown et al, 1984;Anand et al, 1987;Morey et al, 1992;Török, 1995).…”

Quantitative direct probe method for the detection of parvovirus B19

“…In the last decade numerous methods have been used to detect parvovirus B19 in either serum or tissue samples: biotinylated RNA hybridization probes (13); PCR alone or combined with DNA dot hybridization (12,14); strand-specific in situ hybridization probes (15); digoxigenin-labeled DNA hybridization probe (16); in situ hybridization (17); in situ digoxigenin-labeled DNA hybridization probe (18); microplate-capture hybridization of amplified digoxigeninlabeled DNA (19); nested PCR (20); capture hybridization of digoxigenin-labeled B19 DNA amplicons (21), and synthetic oligonucleotides using dot blot (22). We describe a direct hybrid-capture method that is very simple, fast, and does not require expensive equipment.…”

Molecular detection of adeno‐associated virus in cases of spontaneous and intentional human abortion

“…Six-micrometer sections of paraffinized ovular and decidua fragments from all AAV positive cases and from negative cases, for negative control reaction, were mounted on microscope 3-aminopropyl-thrietoxisylane-coated slides. After deparaffination and hydration, nucleic acid was denatured for 2 min in a microwave and ISH was carried out according to Nascimento et al [1991]. The pTAV2 plasmid, with the AAV-2 whole genome inserted, was cloned in Escherichia coli TOP10 (Invitrogen, Life Technologies).…”