Suzuki K, Okamoto N, Kano T
Biomedical Research Center, Olympus Corporation, East Setauket, NY 11733.
J Virol Methods. 1993 Mar;41(3):341-50. doi: 10.1016/0166-0934(93)90023-k.
A rapid and nonradioactive detection method for polymerase chain reaction (PCR) amplified HIV-1 DNA was developed using a colorimetric detection system. Hybridization between biotin-labeled amplified targets and digoxigenin-capture probes occurs in solution followed by efficient and rapid capture onto streptavidin-magnetic beads. The presence of the digoxigenin-capture probe hybridized with biotin-labeled targets is then detected by antidigoxigenin-alkaline phosphatase conjugates using a colorimetric substrate. This approach is highly sensitive and can detect less than 10 HIV targets prior to PCR in approximately 50 min.
利用比色检测系统开发了一种用于聚合酶链反应(PCR)扩增的HIV-1 DNA的快速、非放射性检测方法。生物素标记的扩增靶标与地高辛配基捕获探针在溶液中发生杂交,随后高效快速地捕获到链霉抗生物素蛋白磁珠上。然后使用比色底物,通过抗地高辛配基-碱性磷酸酶偶联物检测与生物素标记靶标杂交的地高辛配基捕获探针的存在。该方法高度灵敏,在PCR之前约50分钟内可检测到少于10个HIV靶标。