Validation Details

Parameter Name: C Reactive Protein (CRP)

Instrument Name: Vitros Fusion 5.1 FS

Validation Period: Dec 2014

Method: Enzymatic Heterogeneous Immunoassay format

Manufacturer: Jhonshon n Jhonshon Vitros

Lab Location: Fortis Mulund

Validation Protocol:

1. Ten Normal specimens were included to verify the reference range.

2. One abnormal sample was assayed 20 times in same run- Intra-assay validation.
3. One abnormal sample was assayed 5 times in 5 different run- Inter-assay variation
4. 5 Samples were parallely assayed with other NABL accredited lab having same test principle-
Interlab correlation
5. Controls provided by the manufacturer- Accuraccy

Results:

1. Normals

S. Sample ID Results (mg/dL)

No
1 0090NJ006755 <5.0
2 0090NJ006768 5.3
3 0090NJ006759 <5.0
4 0090NJ006724 5.1
5 0090NJ006730 9.2
6 0090NJ006731 <5.0
7 0090NJ006728 6.3
8 0090NJ006764 5.2
9 0090NJ006742 <5.0
10 0090NJ006790 <5.0
Mean 6.22
SD 1.73
1.73
2*SD 3.74
12.44
Reference range obtained = Mean±2SD= 2.48-9.96 mg/dL
Manufacturer Reference Range = 0-9.99 mg/dL
2. Intra assay Variation

[Link]. Sample ID CRP

1 PP-1 32.8
2 PP-2 32.6
3 PP-3 32.8
4 PP-4 31.6
5 PP-5 32.7
6 PP-6 33
7 PP-7 33.8
8 PP-8 33.7
9 PP-9 33
10 PP-10 32.4
11 PP-11 32.3
12 PP-12 31.2
13 PP-13 32.3
14 PP-14 32.9
15 PP-15 34.2
16 PP-16 34.7
17 PP-17 33.6
18 PP-18 32.5
19 PP-19 32.3
20 PP-20 32.2
Mean 32.75
SD 0.843
% CV 2.574
3. Inter Assay

[Link]. Sample ID CRP

MOR 1 PPIA-1 63.5
2 PPIA-2 63.8
3 PPIA-3 64.8
4 PPIA-4 63.3
5 PPIA-5 62.2
EVE01 PPIA-6 64
2 PPIA-7 65.8
3 PPIA-8 64.5
4 PPIA-9 62.5
5 PPIA-10 63.4
MOR-02 PPIA-11 65.3
2 PPIA-12 66.8
3 PPIA-13 65
4 PPIA-14 64
5 PPIA-15 65.5
EVE-02 PPIA-16 69
2 PPIA-17 70.9
3 PPIA-18 69.1
4 PPIA-19 68.5
5 PPIA-20 67.6
Mean 64.9
SD 2.42636
% CV 3.73862

4. Linearity
Linearity Range: 5-90 mg/dL

Low Value High Value Results Results %

sample Sample Sample ID
proportion Proportion Expected Obtained Recovery
4 0 6.5 6.5 100
3 1 27.3 29.7 109
2 2 48.1 51.3 107
1 3 68.89 73.3 106
0 4 89.7 89.7 100
Mean 106
 ---

Correlation curve for Linerity CRP

r2 =0.9966
y = 0.96x-0.146
100
90 89.7, 89.7
80
70 68.89, 73.3

60
50 48.1, 51.3
40
30 27.3, 29.7
20
10
6.5, 6.5
0
0 10 20 30 40 50 60 70 80 90 100

5. Interlab correlation

Sample
[Link] Result Interpritation
ID
1 CRP1 76.10 81
2 CRP2 5.30 4.00
3 CRP4 61.50 65.00
4 CRP5 7.50 4.00
5 CRP6 15.90 7.00
6 CRP7 13.90 5.00
7 CRP8 9.30 4.00
 Correlation curve for Inter Lab
comparision CRP
r2 =0.9902 y = 0.96x-0.146
90.00
80.00
81, 76.10
70.00
60.00 65.00, 61.50
50.00
40.00
30.00
20.00
7.00,
5.00, 15.90
13.90
10.00 4.00, 9.30
4.00, 7.50
4.00, 5.30
0.00
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90

Result KDA

6. Accuracy
S. No Controls Expected (mg/dL Obtained (mg/dL)
1 Level 1 19.2 18.08
2 16-22.4 19.2 21.50
3 19.2 19.26
4 19.2 18.04
5 19.2 20.27
6 19.2 21.72
7 19.2 20.28
8 19.2 20.60
9 19.2 21.56
10 19.2 21.29
11 Level 2 63.1 66.25
12 55.1-71.1 63.1 69.50
13 63.1 60.62
14 63.1 62.04
15 63.1 70.84
16 63.1 71.55
17 63.1 69.90
18 63.1 70.58
19 63.1 73.09
20 63.1 65.19
SE=3.14
Control Level 1 = 16-22.4 mg/dL Mean =19.2mg/dL
Control Level 1 = 55.1-71.1mg/dL Mean =63.1mg/dL
 Corelation Curve for Accuracy
CRP
r2=0.9847, Y=
80
60
40 Expected
20 Linear (Expected)
0
0.00 20.00 40.00 60.00 80.00

[Link] Analyte Ref range Intrassay %CV Interassay %CV Linearity Cor Coef
CRP Obse Man Obse Man Obse Man Obse Man Obse Man
mg/dL 2.48-9.96 0-9.99 2.57 3.74 6.5-89.7 5-90 0.9902

Calculation of Systemic error and Total Error using accuracy and precision data

[Link] Analyte %CV Intra %CV Inter Mean SE TE Westgard

1 CRP (mg/dL) 2.57 3.74 3.15 3.14 8.33 NA

Conclusion:

Validation of CRP new kit is successfully completed. Reference range was comparable with the range
recommended by the manufacturer. Inter and Intra assay variation was within the range (%CV <10%)
CRP showed good linearity over the tested range as seen from the linearity data and coefficient of
correlation (r>0.95). Inter lab correlation with other NABL accredited lab working with same principal
showed good correlation (r>0.95). Also the accuracy was found to be acceptable (r>0.95).

The Total error calculated from the precision and accuracy data provides a brief indication of the total
error of the final reported result on this system.

Prepared By: Aarti Jadav Reviewed By: Dr Brijesh Kumar

Approved By: Dr Rajasa Jialdasani