Appendix A
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Note: Although this software and accompanying documentation is dated 2004-2005, it is still valid in 2014. Questions can be sent to CDC-INFO.
Sample Data and Output
A collection of sample data files and program output is included on the distribution diskette. These files may be used to test the program and check results after selecting various options in the different modules. The sample data file is MA112190.DAT and appears as:
| COMMENT:FRASCH ASSAY PB2 1:75 PLATE 1 UNKNOWNS ARE PB2 1:75 | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| 1.716 | 1.757 | 1.769 | 1.840 | 1.462 | 0.960 | 0.562 | 0.313 | 0.171 | 0.090 | 0.041 | 1.440 |
| 1.438 | 1.462 | 1.458 | 1.800 | 1.493 | 1.013 | 0.590 | 0.346 | 0.173 | 0.092 | 0.044 | 1.409 |
| 0.967 | 1.014 | 0.998 | 1.790 | 1.484 | 0.984 | 0.589 | 0.334 | 0.175 | 0.096 | 0.047 | 1.255 |
| 0.586 | 0.606 | 0.627 | 1.801 | 1.472 | 0.970 | 0.588 | 0.322 | 0.169 | 0.085 | 0.043 | 1.239 |
| 0.319 | 0.347 | 0.346 | 1.794 | 1.445 | 0.958 | 0.587 | 0.312 | 0.175 | 0.086 | 0.045 | 0.233 |
| 0.167 | 0.186 | 0.190 | 1.746 | 1.424 | 0.948 | 0.573 | 0.312 | 0.168 | 0.090 | 0.046 | 0.255 |
| 0.094 | 0.103 | 0.099 | 1.791 | 1.531 | 0.955 | 0.584 | 0.325 | 0.170 | 0.091 | 0.040 | 0.001 |
| 0.047 | 0.051 | 0.051 | 1.822 | 1.478 | 0.906 | 0.536 | 0.305 | 0.158 | 0.082 | 0.040 | 0.001 |
| COMMENT:FRASCH ASSAY PB2 1:75 PLATE 2 UNKNOWNS PB2 1:75 | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| 1.830 | 1.843 | 1.914 | 1.898 | 1.637 | 1.045 | 0.620 | 0.341 | 0.178 | 0.091 | 0.046 | 1.446 |
| 1.655 | 1.681 | 1.690 | 1.884 | 1.628 | 1.077 | 0.630 | 0.350 | 0.184 | 0.091 | 0.047 | 1.461 |
| 1.078 | 1.103 | 1.059 | 1.896 | 1.652 | 1.094 | 0.641 | 0.343 | 0.182 | 0.099 | 0.050 | 1.207 |
| 0.655 | 0.658 | 0.635 | 1.898 | 1.644 | 1.091 | 0.623 | 0.348 | 0.173 | 0.095 | 0.048 | 1.203 |
| 0.363 | 0.361 | 0.353 | 1.884 | 1.653 | 1.054 | 0.626 | 0.352 | 0.186 | 0.099 | 0.050 | 0.223 |
| 0.193 | 0.198 | 0.188 | 1.914 | 1.652 | 1.084 | 0.624 | 0.338 | 0.175 | 0.090 | 0.043 | 0.225 |
| 0.091 | 0.095 | 0.094 | 1.841 | 1.612 | 1.079 | 0.624 | 0.349 | 0.172 | 0.068 | 0.046 | 0.001 |
| 0.048 | 0.047 | 0.045 | 1.769 | 1.603 | 1.053 | 0.643 | 0.326 | 0.179 | 0.093 | 0.046 | 0.001 |
The template used to abstract the data is located in file FINAL.TPL. Module 2 may be used to examine the template format and the dilution schedules (refer to Module 2 and Appendix B). Users may run the data abstraction module (Module 3) to produce the .STD, .UNK, .QC, and .LBL files. These are also included on the distribution diskette: MA112190.STD, MA112190.UNK, MA112190.QC, and MA112190.LBL.
Modules 4 may then be used to estimate the four parameters of the logistic-log fit. A sample file of standard curve fitting information is MA112190.FIT contains these results. A portion of that file is reproduced here:
File = C:\ELISAWIN\DOWNLOAD\MA112190.STD- Plate: 1 Project ID = Test Template / Standards ID = Std 1Date: 11/19/96 Time: 11:22:53
FRASCH ASSAY PB2 1:75 PLATE 1 UNKNOWNS ARE PB2 1:75 Results of approximate least squares fitThe procedure used 7 iterations.
Starting values for the iterative process:2.28081 -2.32942 .03409 1.00000
| Dil. | Rel. Dil. | Known O.D. | Pred. O.D. | Residual | % Error | Weight |
|---|---|---|---|---|---|---|
| 0.001666667 | 100.00 | 1.716 | 1.7559 | -0.040 | 2.325 | 1.0000 |
| 0.001666667 | 100.00 | 1.757 | 1.7559 | 0.001 | 0.063 | 1.0000 |
| 0.001666667 | 100.00 | 1.769 | 1.7559 | 0.013 | 0.741 | 1.0000 |
| 0.000833333 | 50.00 | 1.438 | 1.4315 | 0.007 | 0.453 | 1.0000 |
| 0.000833333 | 50.00 | 1.462 | 1.4315 | 0.031 | 2.087 | 1.0000 |
| 0.000833333 | 50.00 | 1.458 | 1.4315 | 0.027 | 1.818 | 1.0000 |
| 0.000416667 | 25.00 | 0.967 | 1.0095 | -0.042 | 4.391 | 1.0000 |
| 0.000416667 | 25.00 | 1.014 | 1.0095 | 0.005 | 0.448 | 1.0000 |
| 0.000416667 | 25.00 | 0.998 | 1.0095 | -0.011 | 1.148 | 1.0000 |
| 0.000208333 | 12.50 | 0.586 | 0.6097 | -0.024 | 4.047 | 1.0000 |
| 0.000208333 | 12.50 | 0.606 | 0.6097 | -0.004 | 0.613 | 1.0000 |
| 0.000208333 | 12.50 | 0.627 | 0.6097 | 0.017 | 2.757 | 1.0000 |
| 0.000104167 | 6.25 | 0.319 | 0.3300 | -0.011 | 3.458 | 1.0000 |
| 0.000104167 | 6.25 | 0.347 | 0.3300 | 0.017 | 4.890 | 1.0000 |
| 0.000104167 | 6.25 | 0.346 | 0.3300 | 0.016 | 4.615 | 1.0000 |
| 0.000052083 | 3.12 | 0.167 | 0.1732 | -0.006 | 3.697 | 1.0000 |
| 0.000052083 | 3.12 | 0.186 | 0.1732 | 0.013 | 6.896 | 1.0000 |
| 0.000052083 | 3.12 | 0.190 | 0.1732 | 0.017 | 8.856 | 1.0000 |
| 0.000026042 | 1.56 | 0.094 | 0.0959 | -0.002 | 2.049 | 1.0000 |
| 0.000026042 | 1.56 | 0.103 | 0.0959 | 0.007 | 6.868 | 1.0000 |
| 0.000026042 | 1.56 | 0.099 | 0.0959 | 0.003 | 3.105 | 1.0000 |
| 0.000013021 | 0.78 | 0.047 | 0.0603 | -0.013 | 28.335 | 1.0000 |
| 0.000013021 | 0.78 | 0.051 | 0.0603 | -0.009 | 18.270 | 1.0000 |
| 0.000013021 | 0.78 | 0.051 | 0.0603 | -0.009 | 18.270 | 1.0000 |
The final coefficients (a, b, c, d) are:
2.13195
-2.10001
0.01841
1.19810
Variances of coefficients =
0.00245
0.00328
0.00000
0.00211
Regular and adjusted sums of squares =
0.007834
0.007834
Approximate multiple R^2 =
0.9991
Logistic-log and spline fit plots may be viewed using Module 5
Module 6 may then be entered to calculate patient serum sample or unknown antibody concentrations. In this sample data set a calibration factor of 1500 was chosen. A complete listing of these results using the unweighted least squares fit is included in file MA112190.RPT. A sample of the report file is reproduced here:
Title:
File: C:\ELISAWIN\DOWNLOAD\MA112190
Project ID = Test Template / Standards ID = Std 1
Date: 11/19/96 Time: 11:23:32
:FRASCH ASSAY PB2 1:75 PLATE 1 UNKNOWNS ARE PB2 1:75
Results from Least Squares fit.
Coefficient file selected: C:\ELISAWIN\DOWNLOAD\MA112190.CFT
| A 2.132 | B -2.100 | C 0.018 | D 1.198 |
Optical densities may range between: 0.032 (l asym) and 2.132 (u asym)
The range of standard dilutions is: 76800 - 600
The range of standard OD's is 0.047 - 1.769
Calibration factor is: 1500
PLATE: 1 UNK# 1 / ID = Unk 1
| # | OD | DILUTION | Avg. OD | CONCENTR. | Avg. CONC. | STD DEV. | CV |
|---|---|---|---|---|---|---|---|
| 01 | 1.840 | 300 | 964.043 | ||||
| 02 | 1.800 | 300 | 1.820 | 850.047 | 907.045 | 80.607 | 8.887 |
| 03 | 1.462 | 600 | 792.541 | ||||
| 04 | 1.493 | 600 | 1.477 | 839.400 | 815.970 | 33.134 | 4.061 |
| 05 | 0.960 | 1200 | 692.807 | ||||
| 06 | 1.013 | 1200 | 0.987 | 754.256 | 723.532 | 43.451 | 6.005 |
| 07 | 0.562 | 2400 | 680.189 | ||||
| 08 | 0.590 | 2400 | 0.576 | 720.796 | 700.493 | 28.713 | 4.099 |
| 09 | 0.313 | 4800 | 708.477 | ||||
| 10 | 0.346 | 4800 | 0.329 | 789.232 | 748.855 | 57.102 | 7.625 |
| 11 | 0.171 | 9600 | 739.668 | ||||
| 12 | 0.173 | 9600 | 0.172 | 749.174 | 744.421 | 6.722 | 0.903 |
| 13 | 0.090 | 19200 | 689.887 | ||||
| 14 | 0.092 | 19200 | 0.091 | 710.248 | 700.068 | 14.398 | 2.057 |
| 15 | 0.041 | 38400 | 286.910 | ||||
| 16 | 0.044 | 38400 | 0.043 | 364.736 | 325.823 | 55.031 | 16.890 |
| Mean (Excludes N/C & Cen): | 708.276 | 166.979 | 23.575 |
PLATE: 1 QUALITY CONTROL
| # | OD | DILUTION | Avg. OD | CONCENTR. | Avg. CONC. | STD DEV. | CV |
|---|---|---|---|---|---|---|---|
| 01 | 1.440 | 1200 | 1523.070 | ||||
| 02 | 1.409 | 1200 | 1.424 | 1441.331 | 1482.200 | 57.798 | 3.899 |
| 03 | 1.255 | 1800 | 1666.735 | ||||
| 04 | 1.239 | 1800 | 1.247 | 1623.825 | 1645.280 | 30.342 | 1.844 |
| 05 | 0.233 | 19200 | 2067.105 | ||||
| 06 | 0.255 | 19200 | 0.244 | 2276.285 | 2171.695 | 147.913 | 6.811 |
| Mean (Excludes N/C & Cen): | 1766.392 | 330.317 | 18.700 |
- Page last reviewed: September 4, 2013
- Page last updated: September 21, 2005
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