Flow cytometry in situ polymerase chain reaction as an innovative approach to HIV-1 proviral DNA detection.
In situ polymerase chain reaction represents an intriguing method to couple the sensitivity of PCR technology with the preservation of cell morphology. The target of this technique is the detection of specific DNA or retrotranscribed RNA inside the cell. In this brief report, we describe an in situ polymerase chain reaction technique revealed by flow cytometry in order to reveal the human immunodeficiency virus type 1 (HIV-1) proviral DNA in peripheral blood mononuclear cells. We also discuss the key steps and parameters of our assay in comparison with current opinion on in situ polymerase chain reaction.
Gibellini
D
D
Department of Clinical and Experimental Medicine, University of Bologna, Italy.
Zauli
G
G
Re
M C
MC
Furlini
G
G
La Placa
M
M
D004279:DNA, Viral / Q000032:analysis*; D005434:Flow Cytometry / Q000379:methods*; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011533:Proviruses / Q000235:genetics / Q000302:isolation & purification
D004279:DNA, Viral