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Qualitative Immunoassay Validation Plan Template
Author: Validation Committee / Document Number: / Equ35-H-02Effective (or Post) Date: / 4 March 2011
Review History / Date of last review: / NA
Reviewed by: / Anne Sholander
SMILE Comments: This document is provided as an example only. It must be revised to accurately reflect your lab’s specific processes and/or specific protocol requirements. Users are directed to countercheck facts when considering their use in other applications. If you have any questions contact SMILE..
Qualitative Immunoassay Validation Plan Template / Document Number / 1610
Effective Date / 1 June 2009
Subject
Template for validation plan of a qualitative Immunoassay method / Page / 1 of 1
Supersedes / Version 1.2
Author(s) / Name, Title / Date
Mark Swartz, Heidi Hanes, Jo Shim, Penny Stevens, Anne Sholander / June 2009
Approved by / Name, Title / Date
SMILE Validation Committee / June 2009
Review History / Date of last review: / 15 February 2011
Reviewed by: / Anne Sholander
Revision History / Version # [0.0] / Revision Date [dd/mm/yy] / Description (notes)
1.1 / 03/12/09 / Added para II.1.b. to indicate that precision is not required unless described by the manufacturer, per DAIDS request.
1.2 / 4/28/10 / Updated within run precision requirements to 20 positive and 20 negative samples.
1.3 / 2/15/2011 / Updated terminology in Diagnostic Sensitivity and Specificity section.
Validation Plan for insert method and test Immunoassay
I. Overview
1. Precision
2. Accuracy
3. Linearity
4. Sensitivity
5. Specificity
6. Reference Range
7. Method Approval
II. Plan: The validation will be conducted on the insert method enzyme immunoassay method. This is a qualitative method.
1. Precision
a. Precision is reproducibility - the agreement of the measurements of replicate runs of the same sample. It is the process of determining the range of random error. The precision is measured in terms of coefficient of variation (CV).
b. Indicate if precision is described by the manufacturer and the methods used. If the manufacturer does not describe precision, indicate in this paragraph that precision is not described by the manufacturer and not applicable to the method.
c. Within run and between run reproducibility will be determined by running the negative control and positive control as follows: For within run, at least 20 replicates of negative control and at least 20 replicates positive control will be tested in one run each. For between run reproducibility, both negative and high positive control will be tested at least once per day but not more than 5 times per day to obtain a total of 20 replicates each.
d. Acceptability criteria: For the between run and within run precision, the optical densities from the negative and positive controls will be used to calculate the coefficient of variance and compared to the manufacturer’s claims for reproducibility. The laboratory CV should be less than or equal to the manufacturer’s stated CV. In the event that an assay does not perform as expected, the Laboratory Director will determine acceptability.
2. Accuracy/Correlation
a. Accuracy is the true value of a substance being measured. Verification of accuracy is the process of determining that the test system is producing true, valid results. This can be verified by:
i. Assaying materials with assigned values
ii. Comparing patient specimen results with a method of long standing use
iii. Verifying results from inter-laboratory survey specimens
iv. Splitting specimens with another sufficiently accredited laboratory.
The results must demonstrate the system is accurate enough to provide clinically valid patient results. Limits of acceptability should be set by the Laboratory Director.
b. Accuracy:
i. Accuracy will also be demonstrated by insert details of accuracy determination
ii. Acceptability criteria: diagnostic sensitivity (true positive rate) and diagnostic specificity (true negative rate) must meet or exceed manufacturer’s stated claims.
3. Linearity/Reportable range
i. Linearity is not applicable to qualitative methods.
ii. The reportable range is negative or positive.
4. Analytical Sensitivity is the lowest concentration of an analyte that can be measured (Lower Limit of Detection). For an FDA approved unmodified method the manufacturer’s stated sensitivity will be used.
5. Analytical Specificity is the determination of the affect of interfering substances. For an FDA approved unmodified method the manufacturer’s stated specificity will be used.
6. Verification of Expected Values/Reference Ranges
Not applicable. Normal (expected) value is insert normal. Abnormal value is insert abnormal.
7. Method Approval
The final decision on methodology validation and acceptance is made after a careful review of all the studies performed as part of the complete method validation process. The Laboratory Director shall make the ultimate decision on method validation. Method acceptance is based on the results from the above studies plus an evaluation of the new method’s cost effectiveness, turn-around-time, laboratory staff training needs, and any other relevant operational considerations.
8. References
a. GCLP Workshop and Workbook18-20 May 2008, Verification of Performance Specifications, pages 1-33.
b. Clinical and Laboratory Standards Institutes (CLSI), User Protocol for Evaluation of Qualitative Test Performance; Approved Guideline--Second Edition. CLSI document EP12-A2 (ISBN 1-56238-654-9). Clinical and Laboratory Standards Institute, 940 West Valley Road, Suite 1400, Wayne, PA 19087-1898 USA, 2008.
c. EP Evaluator Release 8, David G. Rhoads Associates Inc., www.dgrhoads.com.
d. James O. Westgard, Online Validation Training, Westgard QC, Inc. www.westgard.com, Sections 11-Determining Bias,12- Estimating Trueness, and 13- Judging Method Acceptability.
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