To evaluate PVL and CD4+ T-cell decline, peripheral blood was obtained on a weekly and bimonthly basis, respectively. Plasma RNA was extracted utilizing the E.Z.N.A. Viral RNA kit (Omega bio-tek, Norcross, CA) and the manufacturer’s instructions. Viral load was quantified using the iScript One-Step RT-PCR kit with SYBR Green (BioRad, Hercules, CA) per the manufacturer’s instructions as described previously.10 (link),14 (link) CD4+ T-cell decline was determined by staining whole blood with fluorophore conjugated anti-human CD45-FitC (eBioscience), CD3-PE (eBioscience), and CD4-PE/Cy5 (BD Pharmigen, San Jose, CA) antibodies. BD Accuri C6 FACS Analyzer was used to determine cell counts as described previously.13 (link),14 (link) The CD4+ T-cell decline between the infected and uninfected mice was assessed using a two-tailed Student’s t-test (p<0.001).
Protocol detail
Measuring Viral Load and CD4+ T-cell Decline
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Top 5 similar protocols
Protocol cited in 1 other protocol
Variable analysis
independent variables
- Viral load (PVL)
- CD4+ T-cell decline
- Peripheral blood samples collected on a weekly and bimonthly basis for PVL and CD4+ T-cell decline, respectively
- Plasma RNA extraction using the E.Z.N.A. Viral RNA kit
- Viral load quantification using the iScript One-Step RT-PCR kit with SYBR Green
- CD4+ T-cell determination by staining whole blood with fluorophore conjugated anti-human CD45-FitC, CD3-PE, and CD4-PE/Cy5 antibodies and using BD Accuri C6 FACS Analyzer
- Uninfected mice as a control group for CD4+ T-cell decline
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!