Solomon R, Thompson J, Gillespie D
Department of Neoplastic Diseases, Hahnemann University, Philadelphia, Pennsylvania 19102.
J Clin Lab Anal. 1989;3(5):282-6. doi: 10.1002/jcla.1860030505.
Peripheral blood mononuclear cells from HIV-1-infected persons were mixed with 5 M guanidine thiocyanate, and HIV-1-specific probes were hybridized with target RNA directly in the lysate. No RNA purification was needed. Hybrids were purified by repeated capture on superparamagnetic beads coated with oligo(dT) in a device-assisted format, a procedure termed "reversible target capture." Blood mononuclear cells from 70 ARC and AIDS patients were examined and found to have an average of 1.8 x 10(5) molecules of HIV-1 RNA per 2 x 10(6) cells.
将来自HIV-1感染者的外周血单个核细胞与5M硫氰酸胍混合,HIV-1特异性探针直接在裂解物中与靶RNA杂交。无需进行RNA纯化。通过在设备辅助下在涂有寡聚(dT)的超顺磁性珠上反复捕获来纯化杂交体,该过程称为“可逆靶捕获”。对70例艾滋病相关综合征(ARC)和艾滋病患者的血液单个核细胞进行检测,发现每2×10⁶个细胞平均含有1.8×10⁵个HIV-1 RNA分子。