Ghosh M, Kuhn L, West J, Semrau K, Decker D, Thea D, Aldrovandi G; Conference on Retroviruses and Opportunistic Infections.

Abstr 10th Conf Retrovir Oppor Infect Feb 10 14 2003 Hynes Conv Cent Boston Mass USA Conf Retrovir Oppor Infect 10th 2003 Boston Mass. 2003 Feb 10-14; 10: abstract no. 671.

Univ of Alabama at Birmingham

BACKGROUND: The distribution and stability of HIV-1 in breast milk (BM) components remains largely unknown. Inhibitory effects, if any, of BM on HIV RNA and DNA PCR amplification are also poorly understood. We have addressed these issues by using virus-spiked BM samples from HIV- women.METHODS: BM samples from 10 uninfected women were spiked with HIV-1 virions or cells containing a single integrated copy of HIV DNA (8E5/LAV) and incubated under different conditions. Viral RNA was detected by the Roche Amplicor UltraSensitive assay in the whole milk, skim milk, and lipid fractions. HIV-1 DNA was detected by real time polymerase chain reaction assay. Associations between input and observed viral quantity were described using Spearman rank correlation co-efficients (rho), and Kruskall-Wallis tests were used to describe differences in observed viral quantity under different conditions.RESULTS: We found excellent correlation between HIV-1 input copy and recovery in whole milk (rho = 0.965, p < 0.0001), skim milk (rho = 0.972, p < 0.0001), and the lipid fraction (rho = 0.905, p < 0.001). Surprisingly, on average, BM lipid contained over 3 times more (9.38% +/-3.05% to 11.18% +/-2.33%) virus than expected based on the lipid's relative proportion. PCR inhibition was observed in less than 10% of the virion spiked samples. Similar levels of inhibition were noted in a number of BM samples collected from HIV+ women. Detection of HIV proviral DNA in serially diluted 8E5/LAV spiked BM samples also revealed a linear pattern (correlation between the threshold cycles vs log10 DNA copy number > 0.982). The effects of incubation duration and temperature, as well as repeated freeze-thaw cycles on HIV RNA recovery, were also analyzed. It was observed that HIV RNA levels were stable in whole milk, after 3 freeze-thaw cycles and for up to 30 hrs at room temperature.CONCLUSIONS: HIV RNA and DNA can be accurately detected in breast milk. Using a commercial assay to detect HIV RNA, we observed relatively little (

Publication Types:

• Meeting Abstracts

Keywords:

• Acquired Immunodeficiency Syndrome
• Female
• HIV Infections
• HIV Seropositivity
• HIV-1
• Humans
• Milk, Human
• Nucleic Acid Probes
• Polymerase Chain Reaction
• RNA, Viral

Other ID:

• GWAIDS0021679

UI: 102261303

From Meeting Abstracts