%0 Journal Article
%9 ACL : Articles dans des revues avec comité de lecture répertoriées par l'AERES
%A Becquart, Pierre
%A Foulongne, Vincent
%A Willumsen, J.
%A Rouzioux, C.
%A Segondy, Michel
%A Van de Perre, Philippe
%T Quantitation of HIV-1 RNA in breast milk by real time PCR
%D 2006
%L fdi:010035732
%G ENG
%J Journal of Virological Methods
%@ 0166-0934
%K real time PCR ; HIV 1 RNA quantitation ; breast milk
%M CC:0002397693-0017
%N 1
%P 109-111
%R 10.1016/j.jviromet.2005.10.022
%U https://www.documentation.ird.fr/hor/fdi:010035732
%> https://www.documentation.ird.fr/intranet/publi/2006/09/010035732.pdf
%V 133
%W Horizon (IRD)
%X HIV-1 RNA in breast milk is a strong predictor of HIV-1 transmission through breastfeeding. In the present report, breast milk samples from HIV-1 uninfected donors were spiked with dilution of quantified culture supernatant from HIV-1(NDK) infected PBMC. Two RNA extraction techniques based on silica extraction, Nuclisens (R) (BioMerieux) and Triazol (Qiagen), two techniques based on guanidine thiocynanate/chloroforme extraction, TRIzol (Life Technologic) and Amplicor HIV-1 Monitor (TM) (Roche Diagnostic Systems), and one technique based on electrostatic adsorption on iron oxide micro beads (Promega) were compared. HIV-1 RNA was quantitated by real time PCR (LTR gene) and Amplicor HIV-1 Monitor (TM). Combining magnetic micro beads extraction and real time PCR quantitation allowed to correctly quantify breast milk HIV-1 RNA, with a difference between the expected and measured HIV-1 RNA levels always lower than 0.3 log copies/mi. The same combination was confirmed on 25 breast milk samples from HIV-1 infected women collected in Kwazulu-Natal, South Africa, by comparing measurements with those obtained by the Amplicor HIV-1 Monitor (TM) (r(2) = 0.88). Nucleic acid extraction by magnetic micro beads followed by real time PCR is a reliable, sensitive, rapid and simple procedure to quantify HIV-1 RNA in breast milk and allows for PCR inhibitors found frequently in these samples. (c) 2005 Elsevier B.V. All rights reserved.
%$ 052