In situ detection of B19 DNA in bone marrow of immunodeficient patients using a digoxigenin-labelled probe

G. Gentilomi, M. Zerbini, M. Musiani, G. Gallinella, D. Gibellini, S. Venturoli, M. C. Re, S. Pileri, C. Finelli, M. La Placa

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Abstract

An in situ hybridization assay using a digoxigenin-labelled probe was developed to detect B19 DNA in bone marrow erythroid elements of immunodeficient patients with hypoplastic anaemia. A 700 bp Bam HI-Hin dIII fragment of B19 DNA was used to construct the probe by incorporating deoxyuridine triphosphate labelled with digoxigenin. The in situ hybridized B19 DNA probe was visualized by an immunoenzymatic reaction using antidigoxigenin Fab fragments labelled with alkaline phosphatase. Dark blue coloured inclusions at the enzyme site were detected in the nuclei of B19 infected erythroid cells at different stages of cell differentiation. Six out of the nine patients studied showed a positive reaction by in situ hybridization assay. The assay we developed proved highly specific and sensitive and it appears to be a suitable diagnosis test for investigating the possible role of B19 infection as a cause of haematopoietic disorders in immunocompromised hosts.

Original languageEnglish
Pages (from-to)19-24
Number of pages6
JournalMolecular and Cellular Probes
Volume7
Issue number1
DOIs
Publication statusPublished - Feb 1993

Fingerprint

Digoxigenin
In Situ Hybridization
Bone Marrow
Erythroid Cells
Immunoglobulin Fab Fragments
Aplastic Anemia
DNA
DNA Probes
Immunocompromised Host
Alkaline Phosphatase
Cell Differentiation
Enzymes
Infection
deoxyuridine triphosphate

Keywords

  • Parvovirus B19 infection, immunodeficient patients, in situ hybridization

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Gentilomi, G., Zerbini, M., Musiani, M., Gallinella, G., Gibellini, D., Venturoli, S., ... La Placa, M. (1993). In situ detection of B19 DNA in bone marrow of immunodeficient patients using a digoxigenin-labelled probe. Molecular and Cellular Probes, 7(1), 19-24. https://doi.org/10.1006/mcpr.1993.1003

In situ detection of B19 DNA in bone marrow of immunodeficient patients using a digoxigenin-labelled probe. / Gentilomi, G.; Zerbini, M.; Musiani, M.; Gallinella, G.; Gibellini, D.; Venturoli, S.; Re, M. C.; Pileri, S.; Finelli, C.; La Placa, M.

In: Molecular and Cellular Probes, Vol. 7, No. 1, 02.1993, p. 19-24.

Research output: Contribution to journalArticle

Gentilomi, G, Zerbini, M, Musiani, M, Gallinella, G, Gibellini, D, Venturoli, S, Re, MC, Pileri, S, Finelli, C & La Placa, M 1993, 'In situ detection of B19 DNA in bone marrow of immunodeficient patients using a digoxigenin-labelled probe', Molecular and Cellular Probes, vol. 7, no. 1, pp. 19-24. https://doi.org/10.1006/mcpr.1993.1003
Gentilomi, G. ; Zerbini, M. ; Musiani, M. ; Gallinella, G. ; Gibellini, D. ; Venturoli, S. ; Re, M. C. ; Pileri, S. ; Finelli, C. ; La Placa, M. / In situ detection of B19 DNA in bone marrow of immunodeficient patients using a digoxigenin-labelled probe. In: Molecular and Cellular Probes. 1993 ; Vol. 7, No. 1. pp. 19-24.
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