Anti-YFV-Env IgG ELISA Kit

Sample

Serum, Plasma or other Biological Fluids

Species Reactivity

Human

Detection Method

Indirect ELISA

Intended Use

The Human Anti-Yellow Fever Virus Envelope (YFV-ENV) IgG ELISA Kit detects and quantitates YFV envelope protein IgG in human serum or plasma of exposed or immunized animals. This immunoassay is suitable for:
· Determining immune status relative to non-immune controls.
· Assessing efficacy of vaccines, including dosage, adjuvancy, route of immunization and timing.
· Qualifying and/or standardizing vaccine batches and protocols.
This kit is for research use only (RUO), not for diagnosis or therapeutic purposes.

Contents of Kit

1. YFV-ENV Coated Strip Plate 8-well strips (12). Coated with YFV-ENV proteins, post-coated with stabilizers.
2. Anti-YFV-ENV Positive Control 0.65 ml. Anti-YFV-ENV diluted in buffer with protein, detergents and antimicrobial. [Value range on label]
3. Anti-YFV-ENV Calibrator 1 U/ml 0.65 ml
4. Anti-YFV-ENV Calibrator 2.5 U/ml 0.65 ml
5. Anti-YFV-ENV Calibrator 5 U/ml 0.65 ml
6. Anti-YFV-ENV Calibrator 10 U/ml 0.65 ml. Each containing anti-YFV-ENV antibodies; in buffer with antimicrobial.
7. Anti-Human IgG HRP Conjugate (100X) 0.15 ml. Peroxidase conjugated anti-human IgG in buffer with detergents and antimicrobial. Dilute fresh as needed; 10 μl of concentrate to 1 ml of Working Sample/Conjugate Diluent is sufficient for 1 8-well strip. Use within the working day and discard. Return 100X to 2-8°C storage.
8. Sample Diluent (20x) 10 ml. Dilute the entire volume, 10 ml + 190 ml with distilled or deionized water into a clean stock bottle. Label as Working Sample/Conjugate Diluent and store at 2-8°C until the kit lot expires or is used up.
9. Low NSB Sample Diluent 30 ml. Buffer with protein, detergents and antimicrobial. Reduces non-specific binding. Not for HRP Conjugate dilution.
10. Wash Solution Concentrate (100X) 10 ml. Dilute the entire volume 10 ml + 990 ml with distilled or deionized water into a clean stock bottle. Label as Working Wash Solution and store at 4°C for long term and RT for short term.
11. TMB Substrate 12 ml. Chromogenic substrate for HRP containing TMB and peroxide.
12. Stop Solution 12 ml. Dilute sulfuric acid.
13. Product Manual 1 ea.

Storage

The microwell plate and all other reagents, if unopened, are stable at 2-8°C until the expiration date printed on the box label. Stabilities of the working solutions are indicated under Reagent Preparation.

General Description

Yellow fever is a viral hemorrhagic disease spread between humans, as well as between certain other primates and humans, by the bite of yellow fever-infected mosquitoes, primarily of the Aedes aegypti species. The Yellow fever virus (YFV), of the virus family Flaviviridae, is a positive-sense, ssRNA, (11 kb) encoding three structural (C, prM, E) and seven non-structural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B, NS5). Env/E protein is the major inducer and target of virus neutralizing antibodies. The function of the virus E proteins is to attach the virus to receptors on host cells; they initiate the biggest immune response from the host. The M proteins appear to keep the E proteins functional during the assembly of new viruses. C proteins are found in the virus nucleocapsid. YFV vaccine (killed virus) has been very effective and successful. Chimeric variants of YFV, that express E and prM proteins, are being used to produce vaccines against other arbovirus diseases such as dengue fever (DENV), West Nile (WNV), and Japanese encephalitis (JEV).

Vaccines: YFV vaccines, on the WHO's List of Essential Medicines, came into use in 1938. YF-VAX®, Yellow Fever Vaccine, for subcutaneous use, is prepared by culturing the 17D-2048 strain of YFV in chicken embryos. Vaccination may provide life-long immunity, with >90% efficacy, and has been administered to more than 600 million people worldwide. YF-17D is a live-attenuated vaccine that infects and replicates in target cells; recipients of the YF-17D vaccine usually develop detectable viremia after vaccination. Therefore, antibodies to several structural (Env, prM, capsid) and non-structural proteins NS1-5 may be produced. Antibodies to NS1 are also found in natural infection.