A novel approach to nonradioactive hybridization assay of nucleic acids using stained latex particles.
The paper describes a sensitive latex hybridization assay (LHA) method applied for indirect detection of biotinylated nucleic acid hybrids immobilized on a synthetic membrane. The biotinylated hybrids were visualized by means of latex particles containing the fluorescent dye pyronine G and coated with streptavidin; 1.6 and 0.3 pg of lambda-phage DNA was detected by dot blot hybridizations on nylon membrane and polyethyleneimine-cellophane, respectively. The assay sensitivity was increased by three orders of magnitude over that with fluorescently labeled probes due to encapsulation of the fluorescent dye in polymer particles. LHA is simple (single-stage detection procedure), fast, and more sensitive than any of the other nonradioactive hybridization methods.
Vener
T I
TI
Shemyakin Institute of Bioorganic Chemistry, USSR Academy of Sciences, Moscow.
Turchinsky
M F
MF
Knorre
V D
VD
Lukin YuV
Shcherbo
S N
SN
Zubov
V P
VP
Sverdlov
E D
ED
D000171:Acrolein; D001426:Bacterial Proteins; D015415:Biomarkers; D001710:Biotin; D002476:Cellophane; D004247:DNA / Q000737:chemistry*; D005456:Fluorescent Dyes; D007840:Latex; D008567:Membranes, Artificial; D009693:Nucleic Acid Hybridization; D009757:Nylons; D011094:Polyethyleneimine; D011108:Polymers; D013194:Staining and Labeling; D019809:Streptavidin
D001426:Bacterial Proteins; D015415:Biomarkers; D005456:Fluorescent Dyes; D007840:Latex; D008567:Membranes, Artificial; D009757:Nylons; D011108:Polymers; C029975:polyacrolein; D001710:Biotin; D000171:Acrolein; D011094:Polyethyleneimine; D002476:Cellophane; D004247:DNA; D019809:Streptavidin