A new improved method for the concentration of HIV-1 infective particles.
Improvement of the sensitivity of detection systems for human immunodeficiency virus-1 (HIV-1) has been carried out. One approach to improve the sensitivity is purification and/or concentration of the virus from a specimen. In this study, a method for concentrating HIV-1 using polyethylene glycol (PEG) has been re-evaluated and the optimal protocol for concentrating the virus from low-titer specimens was determined. That is, to obtain a virus pellet, a mixture of equal volumes of a specimen and 20% PEG 20,000 solution in saline is incubated at 4 degrees C for 16 h and then centrifuged at 17860 x g in a microcentrifuge for 20 min. HIV-1 in the pellet could be detectable by HIV-1 p24 antigen capture assay for viral protein, reverse transcriptase (RT) assay for viral enzyme, reverse transcriptase polymerase chain reaction (RT-PCR) assay for viral RNA and a virus infectivity assay.
Kohno
Takehiro
T
Department of Microbiology, Osaka Medical College, 2-7 Daigaku-machi, Takatsuki-shi, Osaka 569-8686, Japan.
Mohan
Sharad
S
Goto
Toshiyuki
T
Morita
Chizuko
C
Nakano
Takashi
T
Hong
Wu
W
Sangco
Joseph Carlo E
JC
Morimatsu
Shinichi
S
Sano
Kouichi
K
D002460:Cell Line; D002498:Centrifugation; D015497:HIV-1 / Q000302:isolation & purification*; D006801:Humans; D008828:Microbiological Techniques; D011092:Polyethylene Glycols; D020133:Reverse Transcriptase Polymerase Chain Reaction
D011092:Polyethylene Glycols