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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
1.7161.7571.7691.8401.4620.9600.5620.3130.1710.0900.0411.440
1.4381.4621.4581.8001.4931.0130.5900.3460.1730.0920.0441.409
0.9671.0140.9981.7901.4840.9840.5890.3340.1750.0960.0471.255
0.5860.6060.6271.8011.4720.9700.5880.3220.1690.0850.0431.239
0.3190.3470.3461.7941.4450.9580.5870.3120.1750.0860.0450.233
0.1670.1860.1901.7461.4240.9480.5730.3120.1680.0900.0460.255
0.0940.1030.0991.7911.5310.9550.5840.3250.1700.0910.0400.001
0.0470.0510.0511.8221.4780.9060.5360.3050.1580.0820.0400.001

COMMENT:FRASCH ASSAY PB2 1:75 PLATE 2 UNKNOWNS PB2 1:75
1.8301.8431.9141.8981.6371.0450.6200.3410.1780.0910.0461.446
1.6551.6811.6901.8841.6281.0770.6300.3500.1840.0910.0471.461
1.0781.1031.0591.8961.6521.0940.6410.3430.1820.0990.0501.207
0.6550.6580.6351.8981.6441.0910.6230.3480.1730.0950.0481.203
0.3630.3610.3531.8841.6531.0540.6260.3520.1860.0990.0500.223
0.1930.1980.1881.9141.6521.0840.6240.3380.1750.0900.0430.225
0.0910.0950.0941.8411.6121.0790.6240.3490.1720.0680.0460.001
0.0480.0470.0451.7691.6031.0530.6430.3260.1790.0930.0460.001

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% ErrorWeight
0.001666667100.001.7161.7559-0.0402.3251.0000
0.001666667100.001.7571.75590.0010.0631.0000
0.001666667100.001.7691.75590.0130.7411.0000
0.00083333350.001.4381.43150.0070.4531.0000
0.00083333350.001.4621.43150.0312.0871.0000
0.00083333350.001.4581.43150.0271.8181.0000
0.00041666725.000.9671.0095-0.0424.3911.0000
0.00041666725.001.0141.00950.0050.4481.0000
0.00041666725.000.9981.0095-0.0111.1481.0000
0.00020833312.500.5860.6097-0.0244.0471.0000
0.00020833312.500.6060.6097-0.0040.6131.0000
0.00020833312.500.6270.60970.0172.7571.0000
0.0001041676.250.3190.3300-0.0113.4581.0000
0.0001041676.250.3470.33000.0174.8901.0000
0.0001041676.250.3460.33000.0164.6151.0000
0.0000520833.120.1670.1732-0.0063.6971.0000
0.0000520833.120.1860.17320.0136.8961.0000
0.0000520833.120.1900.17320.0178.8561.0000
0.0000260421.560.0940.0959-0.0022.0491.0000
0.0000260421.560.1030.09590.0076.8681.0000
0.0000260421.560.0990.09590.0033.1051.0000
0.0000130210.780.0470.0603-0.01328.3351.0000
0.0000130210.780.0510.0603-0.00918.2701.0000
0.0000130210.780.0510.0603-0.00918.2701.0000

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

#ODDILUTIONAvg. ODCONCENTR.Avg.
CONC.
STD
DEV.
CV
011.840300 964.043   
021.8003001.820850.047907.04580.6078.887
031.462600 792.541   
041.4936001.477839.400815.97033.1344.061
050.9601200 692.807   
061.01312000.987754.256723.53243.4516.005
070.5622400 680.189   
080.59024000.576720.796700.49328.7134.099
090.3134800 708.477   
100.34648000.329789.232748.85557.1027.625
110.1719600 739.668   
120.17396000.172749.174744.4216.7220.903
130.09019200 689.887   
140.092192000.091710.248700.06814.3982.057
150.04138400 286.910   
160.044384000.043364.736325.82355.03116.890
Mean (Excludes N/C & Cen):   708.276 166.97923.575

 

PLATE: 1 QUALITY CONTROL

#ODDILUTIONAvg. ODCONCENTR.Avg.
CONC.
STD
DEV.
CV
011.4401200 1523.070   
021.40912001.4241441.3311482.20057.7983.899
031.2551800 1666.735   
041.23918001.2471623.8251645.28030.3421.844
050.23319200 2067.105   
060.255192000.2442276.2852171.695147.9136.811
Mean (Excludes N/C & Cen):   1766.392 330.31718.700
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