The HIV Molecular Monitoring Core provides HIV-1 ultrasensitive viral load and sequencing services. In conjunction with the AIDS and Cancer Virus Program Viral Evolution Core, our services are available for studies and research projects from scientists within the National Institutes of Health and the external HIV research community.
The HIV Molecular Monitoring Core works closely with the National Cancer Institute’s HIV Dynamics and Replication Program to coordinate research and testing services involving clinical specimens, with Dr. Frank Maldarelli acting as our clinical specimen liaison.
Measuring impact on HIV-1 viral loads and sequences
We support clinical studies of therapies targeting cancers or other diseases associated with HIV and AIDS. Investigators want to know how the course of treatment impacts the patient’s viral load or sequences, in addition to its effectiveness against the disease in question.
Ultrasensitive HIV-1 assays for plasma, cells, and tissues
Tracking levels of virus is a key means to assess the status of individuals with HIV and to evaluate the impact of treatment interventions in clinical research. We can analyze miniscule levels of HIV, down to 0.15 virions per milliliter of plasma. Participants in clinical studies are typically on antiretroviral treatment, and less-sensitive commercial assays cannot detect their viral loads.
Collaboration Opportunities
We work with investigators through partnership mechanisms orchestrated through FNL's Partnership Development Office.
Our services include: ultrasensitive HIV plasma viral load measurements; cell- and tissue-associated HIV RNA and DNA quantification; and HIV-1 single-genome sequencing and next-gen sequencing analysis for drug-resistance mutations, variant enumeration, and phylogenetic analyses in conjunction with the ACVP Viral Evolution Core.
Email Robert Gorelick with inquiries.
State-of-the-art molecular quantification of HIV
Our work helps keep clinical researchers informed of the impact of an investigational treatment on people living with HIV. The viral load status during treatment trials is often a key endpoint, as investigators need to know whether therapy makes the patient more viremic. In addition to plasma, we evaluate peripheral blood mononuclear cells and tissue samples, including a current study in which we are monitoring viral loads in a person with HIV who underwent a CCR5Δ32 stem cell transplant, which in several rare cases has “cured” a person of HIV-1.
Our capabilities and specializations
Ultrasensitive HIV-1 plasma viral load analysis
HIV-1 is collected from plasma by centrifugation, and nucleic acids are extracted. Then HIV-1 is quantified and reported as HIV-1 copies per milliliter of plasma.
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Nucleic acids for testing isolated essentially quantitatively
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Quantitative RT-PCR analysis using a hybrid real-time/digital approach that provides both high sensitivity and a wide dynamic range
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Assays are highly controlled for HIV-1 DNA contamination and sample inhibition
Cell- and tissue-associated HIV-1 viral load analysis
Nucleic acids are collected from cells and tissues. Then HIV-1 is quantified and reported as HIV-1 copies per 106 cell equivalents. Cell equivalents are assessed from the same DNA sample using an hCCR5 quantitative PCR assay with 2 copies of hCCR5 per diploid cell; this is used to report HIV-1 RNA or DNA copies per 106 cells. All quantitative PCR analyses are performed using a hybrid real-time/digital approach that provides both high sensitivity and a wide dynamic range. Cell- and tissue-associated nucleic acids, both RNA and DNA, are collected nearly quantitatively from the same sample tube using a modified TRIzol extraction protocol.
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Modified TRIzol extraction protocol
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Quantitative RT-PCR analysis (for HIV-1 RNA) and quantitative PCR (for HIV-1 DNA)
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hCCR5 quantitative PCR assay
HIV-1 sequencing services
In conjunction with the AIDS and Cancer Virus Program Viral Evolution Core, sequencing services are provided to clinical investigators for examining HIV-1 drug-resistance mutations, viral evolution, and HIV reservoir studies.
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Single-genome sequencing (Sanger)
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Near-full-length HIV-1 sequencing (Sanger)
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MiSeq™ – next-generation sequencing