Lowering the detection limits of HIV-1 viral load using real-time immuno-PCR for HIV-1 p24 antigen.
Presently, the assay that attains maximal sensitivity and dynamic range of HIV-1 viral copy number (50 copies per milliliter) is nucleic acid amplification of HIV RNA in plasma. Enzyme-linked immunosorbent assay (ELISA) methods for quantification of HIV-1 p24 antigen have been relatively insensitive. In this report, we show data that indicate real-time immuno-polymerase chain reaction (IPCR), a combination of the ELISA and PCR techniques, is more sensitive for HIV-1 p24 antigen detection than other currently reported methods. When derived from an IPCR standard curve, a dose response was observed from patient samples with known viral loads diluted within a 3-log range (1.68-6,514 viral RNA copies per milliliter). IPCR detected 42% (22/52) of patient samples that had fewer than 50 viral RNA copies per milliliter by reverse transcriptase-PCR. IPCR shows the potential to become the most analytically sensitive test available for determination of HIV-1 viral load by the detection of HIV-1 p24 antigen.
Pathology Department, University of Maryland, Baltimore 21201, USA.
Janet M
JM
Barletta
Daniel C
DC
Edelman
Niel T
NT
Constantine
D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D016655:HIV Core Protein p24 / Q000097:blood* / Q000302:isolation & purification; D015658:HIV Infections / Q000175:diagnosis*; D015497:HIV-1 / Q000302:isolation & purification*; D006801:Humans; D012367:RNA, Viral / Q000032:analysis* / Q000097:blood*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D019562:Viral Load
D016655:HIV Core Protein p24; D012367:RNA, Viral