TaqMan RT-PCR and VERSANT HIV-1 RNA 3.0 (bDNA) assay Quantification of HIV-1 RNA viral load in breast milk.
BACKGROUND
Transmission of HIV via breast milk is a primary cause of pediatric HIV infection in developing countries. Reliable methods to detect breast milk viral load are important.


OBJECTIVE
To correlate the ability of the VERSANT HIV 3.0 (bDNA) assay to real-time (RT) TaqMan PCR in quantifying breast milk HIV-1 RNA.


METHODS
Forty-six breast milk samples that had been spiked with cell-free HIV-1 and eight samples spiked with cell-associated HIV-1 were assayed for HIV-1 RNA by both VERSANT HIV 3.0 and TaqMan RNA assays.


RESULTS
Only assays on the cell-free samples were statistically compared. Both a Deming regression slope and a Bland-Altman slope indicated a linear relationship between the two assays. TaqMan quantitations were on average 2.6 times higher than those of HIV 3.0. A linear relationship was observed between serial dilutions of spiked cell-free HIV-1 and both the VERSANT HIV 3.0 and the TaqMan RNA assays.


CONCLUSIONS
The two methods correlated well although the VERSANT HIV 3.0 research protocol quantified HIV-1 RNA slightly lower than TaqMan.
Israel-Ballard
Kiersten
K
Department of Epidemiology, School of Public Health, University of California, 140 Earl Warren Hall, Berkeley, CA 94720-7360, USA. ballardk@berkeley.edu
Ziermann
Rainer
R
Leutenegger
Christian
C
Di Canzio
James
J
Leung
Kimmy
K
Strom
Lynn
L
Abrams
Barbara
B
Chantry
Caroline
C
D021121:Branched DNA Signal Amplification Assay; D015658:HIV Infections / Q000517:prevention & control* / Q000821:virology; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008895:Milk, Human / Q000821:virology*; D012367:RNA, Viral / Q000032:analysis*; D011933:Reagent Kits, Diagnostic; D020133:Reverse Transcriptase Polymerase Chain Reaction
D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic