A plaque hybridization assay for quantifying and cloning infectious human immunodeficiency virus type 1 virions

S. Kato, Y. Hiraishi, N. Nishimura, T. Sugita, M. Tomihama, T. Takano

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

A biological method was developed for quantifying and cloning of infectious virions of human immunodeficiency virus type 1 (HIV-1). Virus preparations were mixed with permissive cells for binding, and the cells were cast in an agarose gel. After incubation for 9 days viral particles released from infected cells propagating from each initially infected cell were transferred on nylon membrane and subjected to hybridization using a radioactive HIV-1 DNA probe. Infectious centers of HIV-1 were detected as hybridization spots on autoradiographs regardless of cytopathic effects or syncytium formation. Three different CD4 + cell lines (MT-4, MOLT-4 and U937) and peripheral blood mononuclear cells from healthy donors were used as recipient cells. Infectious virions were recovered from a portion of agarose gel corresponding to each hybridization spot. This assay is suitable for quantifying infectious HIV-1 virions with different cell tropisms and for investigating the relationship between the phenotype and genotype of HIV-1 at a clonal level.

Original languageEnglish
Pages (from-to)1-7
Number of pages7
JournalJournal of Virological Methods
Volume72
Issue number1
DOIs
Publication statusPublished - 1998 May 1

Keywords

  • HIV-1
  • Infectious virion
  • Plaque hybridization
  • Quantitation

ASJC Scopus subject areas

  • Virology

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