Comparison of Immobilon-N membrane and other membranes for the detection of HIV-1 genome in high risk patients using PCR.
The polymerase chain reaction (PCR) was used for detection of the HIV-1 genome from the peripheral blood lymphocytes of high risk patients. The gag regions of HIV-1 (SK38-SK39) were chosen to amplify viral DNA and the amplified products were spotted onto membrane filters and hybridized with a 32P-labeled SK19 probe. Nitrocellulose, nylon and polyvinylidene difluoride (PVDF) membrane filters were used and compared in dot-blot hybridization. PVDF (Immobilon-N, trade name) filter membranes were demonstrated to be the best membranes on the basis of hybridization data and showed a stronger signal on autoradiograms than the other two types (nitrocellulose and nylon).
Division of Retrovirology and Immunobiology, New York City Dept. of Health, New York 10016.
Y S
YS
Lee
M J
MJ
Collie
L K
LK
Leung
M
M
Seidlin
S T
ST
Beatrice
D003101:Collodion; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D005069:Evaluation Studies as Topic; D005784:Gene Amplification; D005814:Genes, Viral; D015750:Genes, gag; D015497:HIV-1 / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D008567:Membranes, Artificial; D009757:Nylons; D016133:Polymerase Chain Reaction / Q000379:methods*; D011145:Polyvinyls; D014773:Virology / Q000379:methods; D005465:Fluorocarbon Polymers
D003101:Collodion; D004279:DNA, Viral; D008567:Membranes, Artificial; D009757:Nylons; D011145:Polyvinyls; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers