Becquart Pierre, Foulongne Vincent, Willumsen J., Rouzioux C., Segondy Michel, Van de Perre Philippe. (2006). Quantitation of HIV-1 RNA in breast milk by real time PCR. Journal of Virological Methods, 133 (1), p. 109-111. ISSN 0166-0934.
Titre du document
Quantitation of HIV-1 RNA in breast milk by real time PCR
Becquart Pierre, Foulongne Vincent, Willumsen J., Rouzioux C., Segondy Michel, Van de Perre Philippe
Source
Journal of Virological Methods, 2006,
133 (1), p. 109-111 ISSN 0166-0934
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.
