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

Shingo 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

Fingerprint

Virion
HIV-1
Organism Cloning
Sepharose
Gels
Tropism
Nylons
DNA Probes
Giant Cells
Blood Cells
Genotype
Viruses
Phenotype
Cell Line
Membranes

Keywords

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

ASJC Scopus subject areas

  • Virology

Cite this

A plaque hybridization assay for quantifying and cloning infectious human immunodeficiency virus type 1 virions. / Kato, Shingo; Hiraishi, Y.; Nishimura, N.; Sugita, T.; Tomihama, M.; Takano, T.

In: Journal of Virological Methods, Vol. 72, No. 1, 05.1998, p. 1-7.

Research output: Contribution to journalArticle

Kato, Shingo ; Hiraishi, Y. ; Nishimura, N. ; Sugita, T. ; Tomihama, M. ; Takano, T. / A plaque hybridization assay for quantifying and cloning infectious human immunodeficiency virus type 1 virions. In: Journal of Virological Methods. 1998 ; Vol. 72, No. 1. pp. 1-7.
@article{a61534fe9632441f952b12f53cfb5d08,
title = "A plaque hybridization assay for quantifying and cloning infectious human immunodeficiency virus type 1 virions",
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.",
keywords = "HIV-1, Infectious virion, Plaque hybridization, Quantitation",
author = "Shingo Kato and Y. Hiraishi and N. Nishimura and T. Sugita and M. Tomihama and T. Takano",
year = "1998",
month = "5",
doi = "10.1016/S0166-0934(98)00007-X",
language = "English",
volume = "72",
pages = "1--7",
journal = "Journal of Virological Methods",
issn = "0166-0934",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - A plaque hybridization assay for quantifying and cloning infectious human immunodeficiency virus type 1 virions

AU - Kato, Shingo

AU - Hiraishi, Y.

AU - Nishimura, N.

AU - Sugita, T.

AU - Tomihama, M.

AU - Takano, T.

PY - 1998/5

Y1 - 1998/5

N2 - 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.

AB - 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.

KW - HIV-1

KW - Infectious virion

KW - Plaque hybridization

KW - Quantitation

UR - http://www.scopus.com/inward/record.url?scp=0031840868&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031840868&partnerID=8YFLogxK

U2 - 10.1016/S0166-0934(98)00007-X

DO - 10.1016/S0166-0934(98)00007-X

M3 - Article

VL - 72

SP - 1

EP - 7

JO - Journal of Virological Methods

JF - Journal of Virological Methods

SN - 0166-0934

IS - 1

ER -