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 language | English |
|---|---|
| Pages (from-to) | 1-7 |
| Number of pages | 7 |
| Journal | Journal of Virological Methods |
| Volume | 72 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 1998 May |
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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 journal › Article
}
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 -