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CFR (21 CFR part 610.10.e) clearly states that reactive unit should be further tested whenever a supplemental (additional, more specific) test has been approved for such use by the FDA.

Assay Procedure (Manual Assay only)
1.
Carefully remove required number of Strips from the tube and place numbered side up into each well of the Incubation Tray. Include strips for Strong Reactive, Weak Reactive and Non-Reactive controls.
2.
Add 2 mL of Diluted Wash Buffer to each well.
3.
Incubate the strips for at least 5 minutes at room temperature (25 ± 3°C) on a rocking platform (speed of 12 to 16 oscillations per minute). Remove buffer by aspiration.
4.
Add 2 mL of Blotting Buffer to each well.
5.
Add 20 mL each of patients sera or controls to appropriate wells.
6.
Cover the tray with the cover provided and incubate for 1 hour at room temperature (25 ± 3°C) on the rocking platform.
7.
Carefully aspirate the mixture from the wells.
8.
Wash each strip 3 times with 2 mL of Diluted Wash Buffer allowing 5 minutes soak on the rocking platform between each wash.
9.
Add 2 mL of Working Conjugate Solution to each well.
10.
Cover tray and incubate for 1 hour at room temperature (25 ± 3°C) on the rocking platform.
11.
Aspirate Working Conjugate Solution from the wells. Wash as in step 8.
12.
Add 2 mL of Substrate Solution to each well.
13.
Cover tray and incubate for 15 minutes on the rocking platform.
14.
Aspirate the Substrate Solution and rinse the strips at least three times with reagent grade water to stop the reaction.
15.
Using forceps, gently remove strips onto paper towels. Cover with paper towels and dry. Alternatively, allow strips to dry in the wells of the tray.
SUMMARY OF ASSAY PROTOCOLS
Disclaimer Notice
The health information contained herein is provided for educational, reference purposes only, and is not intended to substitute professional medical advice, diagnosis, or treatment with a healthcare provider. All decisions regarding patient care must be made with a healthcare provider, considering the unique characteristics of the patient. Please check the availability and relevance of the information in your respective country.
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