TY - JOUR
T1 - A plaque hybridization assay for quantifying and cloning infectious human immunodeficiency virus type 1 virions
AU - Kato, S.
AU - Hiraishi, Y.
AU - Nishimura, N.
AU - Sugita, T.
AU - Tomihama, M.
AU - Takano, T.
N1 - Funding Information:
We wish to thank Mayako Asakawa for an excellent technical assistance, and Dr. Shigeo Koyasu for critical reading. This work was supported by grants from the Ministry of Education, Science and Culture of Japan and from the Ministry of Health and Welfare of Japan.
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
C2 - 9672127
AN - SCOPUS:0031840868
VL - 72
SP - 1
EP - 7
JO - Journal of Virological Methods
JF - Journal of Virological Methods
SN - 0166-0934
IS - 1
ER -
