Sample Preparation; Principle Of The Procedure; Proper Storage Of Samples And Supplies - Hitachi OPTIGEN AP3600 User Manual

This chapter provides an overview of the patient sera and OPTIGEN assay preparation required before using the AP3600 system.

Principle of the Procedure

The AP3600 OPTIGEN assay is an in vitro test for use in the semi-quantitative determination of circulating allergen-specific IgE
concentrations in human serum. The assay employs a device known as the Pette test chamber to expose patient sera simultaneously to a
number of allergens or allergen mixes.
The AP3600 system automatically performs the following processes:
1. Sample transfer of patient sera from sample vials to sample cups for the assay.
2. Rehydration of the Pette
3. Sample aspiration
4. Sample incubation. Allergen-specific IgE in the serum binds to the allergen-coated wells during the incubation period.
5. Buffer wash. This wash removes unbound serum components.
6. Antibody aspiration
7. Antibody incubation. Enzyme labeled anti-human IgE antibody binds to allergen-specific IgE bound to the allergen-coated wells.
8. Second Buffer wash
9. Photoreagent aspiration
10. Photoreagent incubation. The photoreagent mixture reacts with the labeled antibody to produce chemiluminescence.
11. Measurement of the amount of light emitted by each well. The amount of light is directly proportional to the amount of allergen-
specific IgE in the patient sera.

Proper Storage of Samples and Supplies

The Wash Buffer Concentrate, Antibody, Photoreagents, and Pettes should be stored at 2-8°C. When stored at these temperatures,
the components can be used until the expiration dates printed on the individual component labels.
Sera should be stored at 2-8°C for a period of up to one week. For longer storage, samples should be frozen at -20°C.
Do not freeze kit components.
Note: Repeated freezing and thawing of sera should be avoided. Frozen samples that have been thawed should be thoroughly
mixed prior to centrifugation.
Note: Do not use kit components if signs of deterioration are present. These signs include unusual odor, turbid appearance and
other indications of contamination.

Sample Preparation

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Sample Preparation