Ex vivo explant models are used to characterize in vitro efficacy of preexposure prophylaxis (PrEP) agents. Tissue is challenged with virus in culture and HIV-1 p24 levels are quantified with enzyme-linked immunosorbent assay (ELISA) on supernatants collected throughout a 14-21-day incubation. Due to the narrow dynamic range of HIV-1 p24 kits, we evaluated whether droplet digital PCR (ddPCR) provides an alternative method to quantify HIV-1 replication in supernatant samples. We used samples from the MWRI-01 study, which evaluated the pharmacokinetic/pharmacodynamic profile of long-acting rilpivirine using the explant model (McGowan et al. Lancet HIV 2016). HIV-1 pol RNA was measured with ddPCR, either directly with a one-step method or reverse transcribed to cDNA before ddPCR (two-step method) on supernatants from the MWRI-01 study. Previously analyzed HIV-1 p24 antigen levels (Alliance; Perkin-Elmer) were available for comparison purposes. Both ddPCR methods strongly correlated with HIV-1 p24 and displayed similar patterns of HIV-1 suppression before and after rilpivirine. Compared to the p24 ELISA, two-step and one-step ddPCR reduced the amount of hands-on time by approximately one-half and two-thirds, respectively. ddPCR also required less sample and based on p24 versus ddPCR correlation, could potentially reduce the explant culture time from 14 to 10 days (r = 0.78, p < .001) due to the increased sensitivity of ddPCR. We demonstrate that ddPCR is a suitable alternative to HIV-1 p24 ELISA to quantify HIV-1 infection in the explant model and has the potential to decrease explant culture time.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6434591 | PMC |
| http://dx.doi.org/10.1089/AID.2018.0227 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Background: Due to the unique geographical and climatic conditions in Nagqu (Tibet), the blood station laboratory was only fully established and accredited by 2020. This study validated the performance of the laboratory's blood screening system and analyzed recent trends in blood donation and screening effectiveness.
Methods: Various serum samples were used to assess the performance of hepatitis B, hepatitis C, HIV, and syphilis tests, both serological and nucleic acid tests.
Mitochondrial resilience and antioxidant defence against HIV-1: unveiling the power of extracts and Shatavarin IV.
Front Microbiol
October 2024
Division of Virology, ICMR - National Institute of Translational Virology and AIDS Research, Pune, India.
Article Synopsis
- AR, an Ayurvedic botanical, was studied for its effects on HIV-1 replication and its ability to reduce mitochondrial dysfunction during infection.
- Both AR root extracts and its main active compound, Shatavarin IV, showed non-toxic, dose-dependent inhibition of HIV-1 in laboratory tests, particularly effective against certain viral subtypes.
- Shatavarin IV interacted with key enzymes involved in HIV-1 replication, suggesting its potential as a therapeutic agent, while also helping to alleviate mitochondrial damage caused by the virus.
Efficacy and Possible Mechanism(s) of Action of Gallium Tetraphenylporphyrin Nanoparticles against HIV-TB Coinfection in an Granuloma Structure Model.
ACS Infect Dis
December 2024
Department of Pathology, Microbiology and Immunology, College of Medicine, University of Nebraska Medical Center, Omaha, Nebraska 68198, United States.
Coinfection of (Mtb) and human immunodeficiency virus-1 (HIV) is a significant public health concern. Treatment is challenging due to prolonged duration of therapy and drug interactions between antiretroviral therapy (ART) and anti-TB drugs. Noniron gallium -tetraphenyl porphyrin (GaTP), a heme mimetic, has shown broad antimicrobial activity.
View Article and Find Full Text PDFFourth-Generation HIV Rapid Tests: Enhanced Sensitivity and Reduced Diagnostic Window for HIV-1 Primary Infection Screening.
J Med Virol
November 2024
Laboratoire de Virologie, INSERM, Institut Pierre Louis d' Epidémiologie et de Santé Publique, AP-HP, Hôpitaux Universitaires Pitié Salpêtrière-Charles Foix, Sorbonne Université, Paris, France.
Article Synopsis
- Most HIV rapid tests primarily detect antibodies, which makes them less effective in identifying early HIV infections.
- The Determine test, which includes p24-antigen detection, shows better overall sensitivity compared to the One-Step test, particularly during the early stages of infection.
- Using tests that detect p24-antigen can reduce the diagnostic window significantly, from around 3 months to about 1 month, but caution is advised when testing in the first month after infection.
IFI27 inhibits HIV-1 replication by degrading Gag protein through the ubiquitin-proteasome pathway.
J Virol
November 2024
State Key Laboratory of Genetic Evolution & Animal Models, Key Laboratory of Bioactive Peptides of Yunnan Province, KIZ-CUHK Joint Laboratory of Bioresources and Molecular Research in Common Diseases, Center for Biosafety Mega-Science, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, Yunnan, China.
Article Synopsis
- Type I interferon (IFN-I) and its gene IFI27 are crucial in controlling HIV-1 infection by effectively suppressing viral replication and degrading its proteins, p24 and p55.
- The study found that the IFI27 variant from northern pig-tailed macaques (NPM-IFI27) is particularly effective against HIV-1, utilizing the ubiquitin-proteasome pathway to target and degrade viral proteins.
- These results highlight the potential of NPM-IFI27 as a promising candidate for new antiviral therapies against HIV-1, emphasizing the need for ongoing research in host antiviral agents.
Want AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!