The reagents supplied in this pack are for research use only. 1. Coated microwell strips. Plastic microtitration wells coated with anti-HIV-1 p24 murine monoclonal antibody in foil pouch with desiccant. 1 plate (96 wells) 2. Positive HIV-1 p24 Control 10 ng/ml, 0.1 mL 3. Lysis Buffer, 6 mL 4. Detector Antibody. Anti-HIV-1 p24 conjugated to biotin, 12 mL 5. Conjugate. Streptavidin conjugated to horseradish peroxidase enzyme containing 0.01% Bromonitrodioxane as preservative.12 mL 6. Wash Buffer (20x concentrated). Tris buffered saline pH 7.8-8.0, containing 0.05% Tween 20. Must be diluted before use. 2 Bottles, 60 mL 7. Substrate Solution. Tetramethylbenzidine, 12 mL 8. Stop Solution. 1 N H2SO4, 12 mL
Storage
All reagents should be stored at 2-8°C and should not be used beyond the expiration date on the label. Once opened, microtitration strips may be stored at 2-8°C until the expiration date on the label, provided that desiccated conditions are maintained. Unused strips should be returned to their original foil pouch along with the sachet of desiccant. Secure open foil pouch using zip top before storage. The working strength Wash Buffer should not be stored for longer than 3 weeks at 2-8°C. It is recommended that Wash Buffer be freshly diluted before each assay. If the working strength buffer becomes visibly cloudy or develops precipitate during the 3 weeks, do not use it. Indications of Deterioration The HIV-1 p24 Assay may be considered to have deteriorated if: 1. The kit fails to meet the required criteria for a valid test (see Interpretation of Results). 2. Reagents becoming visibly cloudy or develop precipitate. Note: Concentrated Wash buffer, when cold, normally develops crystalline precipitates, which re-dissolve on heating at 37°C. 3. The Substrate Solution turns dark blue. This is likely to be caused by chemical contamination of the Substrate Solution.
Precision
Four samples with different levels of activity were assayed ten times each on three different assays. The results are summarized in the following table.
Sensitivity
To determine the sensitivity of the assay, the 0 standard was assayed 20 times. The minimal detectable level was calculated by adding two standard deviations to the mean absorbance for the 0 standard. The minimal detectable level is 1.7 pg/mL.
Citations
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Background
Acquired immunodeficiency syndrome (AIDS) is a disease spread by the human immunodeficiency virus (HIV) that threatens to take human life. It is a retrovirus, and on the basis of genome architecture and sequence similarity HIV could be separated into HIV-1 and HIV-2. The leading cause of the global AIDS pandemic is HIV-1, and HIV-2 is a limited epidemic mainly in West Africa, with low virulence and transmission and protracted course. Then we have the four subtypes of HIV-1 that are also homologous in gene sequence, M, N, O and P, where strains of group M circulate globally. HIV-1 is highly mutagenic and highly recombinant: there will always be circulating recombinant forms (CRFs) and unique recombinant forms (URFs). HIV-1 subtype variation is a manifestation of how the virus evolved in different geographical areas and groups of people, and genetic variations between strains can affect viral spread, progression of disease, drug treatment and vaccine development.
Figure 1. Distribution of HIV-1 Group M subtypes and CRF families (Source: Torrecilla E, et al. 2014)
There are several factors that support HIV-1 genetic diversity and patterns of rapid evolution. It's not only HIV-1 genes that display such genetic diversity and fast evolutionary cycles. The HIV-1 reverse transcriptase doesn't have a corrective role, and so HIV-1 is very susceptible to mutations at replication time. On average, each round of replication produces about 0.2 base variants per genome, including base substitutions, insertions or deletions, and recombinations. The relatively small genome of HIV-1 and its rapid replication efficiency result in the production of 1010/1012 new virions per day within an infected individual. In addition, host immune selection pressure and antiviral drug screening pressure have accelerated the evolution of HIV-1 mutations. Studies on the specific humoral immune response mediated by HIV-1 antibodies have shown that neutralizing antibodies play a key role in protecting the host from viral infection and slowing disease progression. Neutralizing antibodies specifically recognize and bind to the glycoprotein on the surface of the enveloped virus, thereby preventing the protein from mediating viral adsorption to target cells and fusion with the cell membrane. However, neutralizing antibody-mediated immune selection pressure predisposes to the generation of HIV-1 escape mutations.
Highly active anti-retroviral therapy (HAART) is now the standard therapy in both AIDS and HIV-1 patients, and significantly prolongs life and slows down disease progression. But because HIV is late, there is still a major hurdle to getting rid of the virus entirely and getting a cure in treatment. Researchers have proposed a functional cure, which involves maintaining normal body immune function and long-term viral suppression without receiving or interrupting HAART.
Alternative Names
Human immunodeficiency virus 1 p24 ELISA Kit Human immunodeficiency virus type 1 p24 ELISA Kit
References
1. Torrecilla E, et al. New findings in cleavage sites variability across groups, subtypes and recombinants of human immunodeficiency virus type 1. PLoS One. 2014 Feb 7;9(2):e88099.
2. Arenas M. Genetic Consequences of Antiviral Therapy on HIV-1. Comput Math Methods Med. 2015;2015:395826.
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References
Integrated Pharmacy and PrEP Navigation Services to Support PrEP Uptake: A Quality Improvement Project
JANAC-JOURNAL OF THE ASSOCIATION OF NURSES IN AIDS CARE
Preexposure prophylaxis (PrEP) is highly effective in preventing HIV among both men and women, with the reduction in risk directly linked to medication adherence. Navigation services and other adherence interventions have demonstrated efficacy in medication uptake; however, their use may not be fully integrated into clinic operations or their roles clearly defined. This quality improvement (QI) project developed an evidenced-based PrEP Navigation (PN) tool to identify patient-reported barriers to uptake and to support process improvement at a large community health center in Washington, DC. Outcomes related to patient-reported barriers, patient demographics, and time to medication pickup from the pharmacy were measured before and after implementation. A total of 198 patients were included in this analysis. Mean days from initial prescription to medication pickup was reduced by 1.42 days (p = .030) following PN tool implementation. The evidenced-based PN tool is modifiable to the needs of the individual clinic and the patients they care for to support wide-scale PrEP uptake and continuous system process improvements.
E-cigarette Perceptions among HIV-positive Smokers in a Switching Study
TOBACCO REGULATORY SCIENCE
Authors: Cioe, Patricia A.; Tidey, Jennifer W.; Mercurio, Alana N.; Costantino, Catherine; Kahler, Christopher W.
Objectives: Cigarette smoking is highly prevalent among persons with HIV (PWH), which is concerning due to their increased risk of developing smoking-related cancers compared to those without HIV. Smoking cessation rates are substantially lower among PWH compared to the general population and less than half report a goal of total abstinence. Given these factors, a switch to electronic cigarettes (ECs), as a form of tobacco harm reduction, may be a viable approach to decreasing tobacco-related morbidity and mortality in PWH. The US Food and Drug Administration has the authority to implement product standards for cigarettes, including ECs and flavors. Methods: This qualitative study enrolled 19 HIV-positive smokers into an EC switch study. At Week 12, qualitative interviews were conducted to examine EC perceptions and flavor preferences. Results: ECs were viewed as a less harmful, cost-effective method of reducing or eliminating CC smoking, and non-tobacco flavors were an essential part of EC appeal and use. Conclusions: Flavored EC liquid seems to enhance the user experience and may influence the user's ability to make a complete switch. Tobacco harm reduction, as a strategy, will only be effective if current evidence guides tobacco regulatory decisions.
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Figure 1. Distribution of HIV-1 Group M subtypes and CRF families
