Detection of HIV-1 DNA in pediatric AIDS brain tissue by two-step ISPCR.
In order to detect latent infection in neurons or other cell types in formalin-fixed brain tissue, we performed polymerase chain reaction amplification with incorporation of digoxigenin-conjugated deoxynucleotides, followed by in situ hybridization with biotinylated probes. The use of this two-step technique in brain tissue from a child with severe HIV-1 encephalitis revealed signal in both nuclear and perinuclear regions of cells identified as monocytes and astrocytes, and also in perineuronal satellite cells of glial morphology, but HIV-1 infection of neurons was not detected.
Sharer
L R
LR
Department of Pathology, New Jersey Medical School, Newark.
Saito
Y
Y
Epstein
L G
LG
Blumberg
B M
BM
D015526:AIDS Dementia Complex / Q000473:pathology / Q000821:virology; D000163:Acquired Immunodeficiency Syndrome / Q000473:pathology / Q000821:virology*; D001921:Brain / Q000473:pathology / Q000821:virology*; D002648:Child; D004279:DNA, Viral / Q000302:isolation & purification*; D004076:Digoxigenin; D018792:Encephalitis, Viral / Q000473:pathology / Q000821:virology*; D015497:HIV-1 / Q000302:isolation & purification*; D006801:Humans; D017403:In Situ Hybridization; D008297:Male; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D011533:Proviruses / Q000302:isolation & purification*; D016707:Tissue Fixation
D004279:DNA, Viral; D004076:Digoxigenin