Clinical Chemistry / SEVEN CA 125 IMMUNOASSAYS

Performance Characteristics of Seven Automated CA 125 Assays

Shella K. Mongia, MD,1 Mindy L. Rawlins,2 William E. Owen, MT(ASCP),2 and William L. Roberts, MD, PhD1
Key Words: CA 125; Immunoassay; Imprecision; Linearity; Method comparison; Ovarian carcinoma; Tumor marker
DOI: 10.1309/NBA312W0LANRXYH9

Abstract
Cancer antigen 125 (CA 125) is a high-molecularmass glycoprotein that is used as a tumor marker to monitor disease progression and response to therapy and in early detection of recurrence after treatment for ovarian cancer. The Access 2 (Beckman Coulter, Brea, CA), ADVIA Centaur (Bayer Diagnostics, Tarrytown, NY), ARCHITECT i2000 (Abbott Diagnostics, Abbott Park, IL), AxSYM (Abbott Diagnostics), Elecsys 2010 (Roche Diagnostics, Indianapolis, IN), IMMULITE 2000 (Diagnostic Products, Los Angeles, CA), and VITROS ECi (Ortho Clinical Diagnostics, Raritan, NJ) assays for CA 125 were evaluated for detection limit, dilution linearity, imprecision, correlation, and reference intervals. The maximum average deviation from target recoveries for dilution linearity studies ranged from 3.7% for the ADVIA Centaur to 18.2% for the IMMULITE 2000. Imprecision studies yielded total coefficients of variation of 2.0% to 8.3% at CA 125 concentrations of 35 and 114 U/mL (35 and 114 kU/L). Method comparison studies revealed good agreement with the VITROS ECi comparison method, with slopes ranging between 0.88 and 1.19 and correlation coefficients of more than 0.95. All methods show acceptable performance characteristics and generally compare well. However, for some samples, substantial differences exist between methods, necessitating parallel testing when introducing a new method.

Cancer antigen 125 (CA 125) is a high-molecular-mass glycoprotein (>200 kd) and initially was recognized by monoclonal antibody OC125. The OC125 monoclonal antibody was generated by immunization of BALB/c mice with the OVCA43 cell line isolated from ascites fluid of a patient with serous papillary cystadenocarcinoma of the ovary.1 CA 125 carries 2 major antigenic domains, and monoclonal antibodies against it can be classified as OC125-like or M11like.2 CA 125 contains 24% carbohydrate and is expressed normally in fetal coelomic epithelium, by epithelial ovarian tumors, normal and pathologic tissues of mllerian duct origin, and mesothelial cells.3,4 The CA 125 assay is useful for determining the prognosis of endometrial carcinoma, in evaluation of advanced endometriosis, and as an aid in monitoring the response to therapy for patients with epithelial ovarian cancer.5 The assay is not useful for screening for ovarian cancer in asymptomatic women.6 An immunoradiometric assay for CA 125 was first commercialized by Centocor (now Fujirebio Diagnostics, Malvern, PA). That assay used the OC125 monoclonal antibody for capture and tracer functions. To improve on the sensitivity and specificity of the first-generation assays, a double-determinant monoclonal antibody based format was developed using 2 monoclonal antibodies with specificities against the antigenic domains OC125 and M11. The second-generation assays have been reported to show better agreement than the first-generation assays.7-13 In the present study, 7 automated second-generation immunoassays for CA 125 were evaluated for detection limit, dilution linearity, imprecision, method comparison, and reference intervals.
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Materials and Methods

The 7 assays that were evaluated were the Access 2 (Beckman Coulter, Brea, CA), ADVIA Centaur (Bayer Diagnostics, Tarrytown, NY), ARCHITECT i2000 (Abbott Diagnostics, Abbott Park, IL), AxSYM (Abbott), Elecsys 2010 (Roche Diagnostics, Indianapolis, IN), IMMULITE 2000 (Diagnostic Products, Los Angeles, CA), and VITROS ECi (Ortho Clinical Diagnostics, Raritan, NJ). All assays were performed in 1 laboratory according to the manufacturers instructions. The limit of detection of each method was determined by analyzing 10 replicates of the zero calibrator and 4 replicates of the lowest nonzero calibrator (15-35 U/mL [15-35 kU/L]) from each manufacturer. The data were analyzed using EP Evaluator Release 5 software (David G. Rhoads Associates, Kennett Square, PA). The results of 4 separate runs were averaged. For the Access 2, the Cal S0 (0 U/mL [0 kU/L]) and Calibrator S1 (25 U/mL [25 kU/L]) were used. For the ADVIA Centaur, CA 125 MCM1 zero and Calibrator 1 (16.4 U/mL [16.4 kU/L]) were used. For the ARCHITECT i2000, CA 125 Calibrator A (0 U/mL [0 kU/L]) and Calibrator B (20 U/mL [20 kU/L]) were used. For the AxSYM, CA 125 Calibrator A (0 U/mL [0 kU/L]) and Calibrator B (15 U/mL [15 kU/L]) were used. For the Elecsys 2010, CA 125 CalCheck1 (0 U/mL [0 kU/L]) and Calibrator 1 (35 U/mL [35 kU/L]) were used. For the IMMULITE 2000, the IMMULITE OV-MA diluent (0 U/mL [0 kU/L]) and Low Adjustor (16 U/mL [16 kU/L]) were used. For the VITROS ECi, sample diluent B (0 U/mL [0 kU/L]) and CA 125 Calibrator 1 (24 U/mL [24 kU/L]) were used. Dilution linearity was assessed by pooling 2 or 3 patient samples with similar high CA 125 concentrations for each method, making serial dilutions (1:2 to 1:32), and analyzing each sample in triplicate. The upper limits chosen were based on the analytic measurement range stated in each assays package insert. For the Access 2 and Elecsys 2010 methods, the concentration of the high pool was approximately 5,000 U/mL (5,000 kU/L); for the ARCHITECT i2000 and VITROS ECi, the high pool was approximately 1,000 U/mL (1,000 kU/L); and for the ADVIA Centaur, AxSYM, and IMMULITE 2000 methods, the high pool was approximately 500 U/mL (500 kU/L). Whenever appropriate, the pooled patient samples were first diluted with the manufacturers recommended diluent until they were within the analytic measurement range of each method. For the Access 2, the patient sample with high CA 125 values was serially diluted with Diluent A (Beckman Coulter) to give final concentrations of 0.3%, 0.78%, 1.5%, 3.1%, 6.25%, 12.5%, 25%, 50%, and 100% (range of CA 125 concentrations, 23-5,087 U/mL [23-5,087 kU/L]). For the ADVIA Centaur, serial dilutions were made with MD1 diluent (Bayer
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Diagnostics) to give final concentrations of 1.5%, 3.1%, 6.25%, 12.5%, 25%, 50%, and 90% (range of CA 125 concentrations, 10-538 U/mL [10-538 kU/L]). For the ARCHITECT i2000, serial dilutions were made with ARCHITECT wash buffer (Abbott Diagnostics) to give final concentrations of 3.1%, 6.25%, 12.5%, 25%, 50%, and 90% (range of CA 125 concentrations, 37-960 U/mL [37-960 kU/L]). For the AxSYM, serial dilutions were made with AxSYM CA 125 diluent (Abbott Diagnostics) to give final concentrations of 3.1%, 6.25%, 12.5%, 25%, 50%, and 90% (range of CA 125 concentrations, 17-494 U/mL [17-494 kU/L]). For the Elecsys 2010, linearity was assessed by serial dilution with Universal diluent (Roche Diagnostics) to give final concentrations of 0.39%, 0.78%, 1.5%, 3.1%, 6.25%, 12.5%, 25%, 50%, and 100% (range of CA 125 concentrations, 22-4,720 U/mL [22-4,720 kU/L]). For the IMMULITE 2000, the samples were diluted with OV diluent (Diagnostic Products) to give final concentrations of 2.25%, 4.5%, 9%, 18%, 36%, 73%, 81%, and 90% (range of CA 125 concentrations, 16-484 U/mL [16-484 kU/L]). For the VITROS ECi, linearity was assessed by diluting the sample with High sample diluent B (Ortho Clinical Diagnostics) to obtain final concentrations of 1.5%, 3.1%, 6.25%, 12.5%, 25%, 50%, and 100% (range of CA 125 concentrations, 13.5-975 U/mL [13.5-975 kU/L]). Assessment of dilution linearity was performed with EP Evaluator Release 5 software. Imprecision studies were performed by assaying Lyphocheck Tumor Marker Controls (Bio-Rad Laboratories, Hercules, CA) and the manufacturers high control for each analyzer, analyzed in replicates of 2 per run. Two runs per day were conducted on each of 5 days with a minimum of 2 hours separating each run for the total of 20 replicates for each level of quality control material. Assay imprecision data were analyzed by using EP Evaluator Release 5 software. Correlation studies were performed with 98 specimens from females between the ages of 14 and 87 years and with CA 125 concentrations from 0 to 1,000 U/mL (0-1,000 kU/L). The samples were tested according to the package insert instructions for each method. The samples with measured concentrations of more than the analytic measurement range for each method were rerun after dilution according to the manufacturers instructions. The VITROS ECi was chosen as the comparison method because it correlated the best with all of the other methods. Passing-Bablok regression analysis was performed using Analyse-it + Clinical Laboratory, version 1.63 software (Analyse-It Software, Leeds, England). Reference intervals were determined using samples from 120 apparently healthy women between 20 and 65 years of age who were not taking any prescription medications. One subject with the highest CA 125 concentration in this group was found to have ovarian cancer; therefore, her samples were excluded from further analysis. Statistical analysis was performed using
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EP Evaluator Release 5 software. The 97.5 percentile upper reference limit was determined using a transformed parametric approach. All studies using samples from human subjects were approved by the institutional review board of the University of Utah, Salt Lake City.

Results
The limit of detection ranged from 0.05 U/mL (0.05 kU/L) on the Access 2 to 1.45 U/mL (1.45 kU/L) on the AxSYM. The limit of detection was 0.30 U/mL (0.30 kU/L) for the ADVIA Centaur method, 0.35 U/mL (0.35 kU/L) for the ARCHITECT i2000 method, 0.09 U/mL (0.09 kU/L) for the Elecsys 2010 method, 0.10 U/mL (0.10 kU/L) for the IMMULITE 2000 method, and 0.06 U/mL (0.06 kU/L) for the VITROS ECi method. In all cases, the detection limit we measured was lower than the manufacturers claim. For dilution linearity studies, the maximum average deviation from target recovery was 10.9% for the Access 2 at 162 U/mL (162 kU/L), 3.7% for the ADVIA Centaur at 150 U/mL (150 kU/L), 8.2% for the ARCHITECT i2000 at 128 U/mL (128 kU/L), 7.7% for the AxSYM at 35 U/mL (35 kU/L), 10.7% for the Elecsys 2010 at 151 U/mL (151 kU/L), 18.2% for the IMMULITE 2000 at 52 U/mL (52 kU/L), and 4.1% for the VITROS ECi at 120 U/mL (120 kU/L). It is noteworthy that the upper limit of the analytic measurement range was 5,000 U/mL (5,000 kU/L) for the Access 2 and Elecsys 2010
Table 1 Summary of Imprecision Data*

methods, 1,000 U/mL (1,000 kU/L) for the ARCHITECT i2000 and VITROS ECi methods, 600 U/mL (600 kU/L) for the ADVIA Centaur method, 500 U/mL (500 kU/L) for the AxSYM method, and 400 U/mL (400 kU/L) for the IMMULITE 2000 method based on our studies with 1 lot of reagent. Approximately 0.5% of the CA 125 results generated in our laboratory are greater than 5,000 U/mL (5,000 kU/L), 3.2% are greater than 1,000 U/mL (1,000 kU/L), and 5.6% are greater than 500 U/mL (500 kU/L). The imprecision of each assay was assessed by using commercial serum-based quality control materials. The coefficients of variation (CVs) for the low control (35 U/mL [35 kU/L]) were 1.3% to 8.3% within run, 0% to 3.1% between runs, and 2.0% to 8.3% total Table 1. For the high control (114 U/mL [114 kU/L]), CVs were 1.4% to 4.3% within run, 0% to 3.2% between runs, and 1.8% to 6.3% total. Results from method comparison studies with PassingBablok analysis are shown Figure 1. Slopes ranged from 0.87 for the IMMULITE 2000 assay to 1.19 for the ARCHITECT i2000. Difference plots for the same data are shown Figure 2. The mean differences ranged from 30 U/mL (30 kU/L) for IMMULITE 2000 to 63 U/mL (63 kU/L) for the ARCHITECT i2000 method. The IMMULITE 2000 had 2 results that were lower than the VITROS ECi method by 250 U/mL (250 kU/L) or more (Figure 2F). The analytic concordance of each method with the comparison method for these samples is shown using the manufacturers suggested upper reference limits Table 2.

Coefficient of Variation (%) Assay Access 2 ADVIA Centaur ARCHITECT i 2000 Control Material Ly1 Ly2 Ly1 Ly2 Mfg3 Ly1 Ly2 Mfg2 Mfg3 Ly1 Ly2 Mfg3 Ly1 Ly2 Ly1 Ly2 Mfg3 Ly1 Ly2 Mfg3 Mean Concentration (U/mL) 29.6 103 39.7 130 237 43.3 135 306 656 42.2 136 208 34.1 101 28.2 99.6 126 27 .9 95.7 267 Within-Run 1.4 1.5 2.5 3.1 2.2 1.7 2.5 2.5 1.6 8.3 4.3 7 .1 2.0 1.4 4.5 2.9 4.1 1.3 2.1 1.3 Between-Run 0.9 1.4 2.3 2.2 0.0 3.1 0.0 0.4 0.0 0.0 3.2 0.0 0.0 0.9 0.0 0.0 0.0 1.1 0.6 0.6 Total 2.0 3.0 3.3 3.8 2.4 3.5 2.9 2.6 1.6 8.3 6.3 7 .1 2.4 1.8 4.5 3.7 4.3 2.1 2.2 1.4

AxSYM Elecsys 2010 IMMULITE 2000 VITROS ECi

Ly, Lyphocheck control material (Bio-Rad Laboratories, Hercules, CA); Mfg, control material from the method manufacturer. * Values are given in conventional units; to convert to Systme International units (kU/L), multiply by 1. Access 2, Beckman Coulter, Brea, CA; ADVIA Centaur, Bayer Diagnostics, Tarrytown, NY; ARCHITECT i2000, Abbott Diagnostics, Abbott Park, IL; AxSYM, Abbott Diagnostics; Elecsys 2010, Roche Diagnostics, Indianapolis, IN; IMMULITE 2000, Diagnostic Products, Los Angeles, CA; VITROS ECi, Ortho Clinical Diagnostics, Raritan, NJ.

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ARCHITECT i 2000 CA 125 (U/mL)

ADVIA Centaur CA 125 (U/mL)

1,250 Access 2 CA 125 (U/mL) 1,000 750 500 250 0 0 250 500 750 1,000 1,250 VITROS ECi CA 125 (U/mL)

1,250 1,000 750 500 250 0 0 250 500 750 1,000 1,250 VITROS ECi CA 125 (U/mL)

1,250 1,000 750 500 250 0 0 250 500 750 1,000 1,250 VITROS ECi CA 125 (U/mL)

D
1,250 AxSYM CA 125 (U/mL) 1,000 750 500 250 0 0 250 500 750 1,000 1,250 VITROS ECi CA 125 (U/mL)

E
1,250 1,000 750 500 250 0 0 250 500 750 1,000 1,250 VITROS ECi CA 125 (U/mL)

F
1,250 1,000 750 500 250 0 0 250 500 750 1,000 1,250 VITROS ECi CA 125 (U/mL)

Figure 1 Comparison of CA 125 methods by Passing-Bablok analysis. The solid line indicates the Passing-Bablok regression line, and the dashed line indicates y = x. Data for 98 samples are shown in each panel. A, Slope, 1.12 (95% confidence interval [CI], 1.11 to 1.51); intercept, 1.14 (95% CI, 0.021 to 2.73); r = 0.98. B, Slope, 0.96 (95% CI, 0.93 to 0.99); intercept, 2.4 (95% CI, 0.84 to 4.27); r = 0.98. C, Slope, 1.19 (95% CI, 1.7 to 1.21); intercept, 5.37 (95% CI, 3.63 to 7 .94); r = 0.99. D, Slope, 1.05 (95% CI, 1.03 to 1.1); intercept, 1.83 (95% CI, 4.54 to 0.46); r = 0.98. E, Slope, 0.95 (95% CI, 0.92 to 0.98); intercept, 12.07 (95% CI, 8.79 to 15.78); r = 0.97 . F, Slope, 0.87 (95% CI, 0.84 to 0.92); intercept, 1.48 (95% CI, 5.84 to 0.72); r = 0.95. Access 2, Beckman Coulter, Brea, CA; ADVIA Centaur, Bayer Diagnostics, Tarrytown, NY; ARCHITECT i 2000, Abbott Diagnostics, Abbott Park, IL; AxSYM, Abbott Diagnostics; Elecsys 2010, Roche Diagnostics, Indianapolis, IN; IMMULITE 2000, Diagnostic Products, Los Angeles, CA; VITROS ECi, Ortho Clinical Diagnostics, Raritan, NJ.

Reference interval information obtained using samples from apparently healthy women is shown Table 3. The median CA 125 concentration varied from 6.3 U/mL (6.3 kU/L) for the IMMULITE 2000 to 17.2 U/mL (17.2 kU/L) for the ELECSYS 2010 method.

Discussion
The detection limit of all 7 immunoassays was less than 1.5 U/mL (<1.5 kU/L) and was lower than manufacturers
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claimed sensitivity. Linearity was acceptable for all methods, with recoveries within 10% of the target values except for the IMMULITE 2000 method, in which an 18.2% deviation from the target value was observed. The manufacturers claimed analytic measurement range for the IMMULITE 2000 is up to 500 U/mL (500 kU/L), but our data for 1 lot of reagent demonstrated linearity only to 400 U/mL (400 kU/L). The imprecision of all automated assays was adequate, with total CVs of less than 8.3% even at CA 125 concentrations that were at or near a clinical decision threshold of 35 U/mL (35 kU/L).
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IMMULITE 2000 CA 125 (U/mL)

Elecsys 2010 CA 125 (U/mL)

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B
ARCHITECT i 2000 VITROS ECi CA 125 (U/mL)

ADVIA Centaur VITROS ECi CA 125 (U/mL)

Access 2 VITROS ECi CA 125 (U/mL)

500 250 0 250 500 0 250 500 750 1,000 VITROS ECi CA 125 (U/mL)

500 250 0 250 500 0 250 500 750 1,000 VITROS ECi CA 125 (U/mL)

500 250 0 250 500 0 250 500 750 1,000 VITROS ECi CA 125 (U/mL)

E
IMMULITE 2000 VITROS ECi CA 125 (U/mL) Elecsys 2010 VITROS ECi CA 125 (U/mL)

AxSYM VITROS ECi CA 125 (U/mL)

500 250 0 250 500 0 250 500 750 1,000 VITROS ECi CA 125 (U/mL)

500 250 0 250 500 0 250 500 750 1,000 VITROS ECi CA 125 (U/mL)

500 250 0 250 500 0 250 500 750 1,000 VITROS ECi CA 125 (U/mL)

Figure 2 Comparison of CA 125 methods using difference plots. The solid line indicates the mean difference between methods, and the dashed lines indicate the upper and lower 95% confidence limits of the difference between methods. Data for 98 samples are shown in each panel. A, Mean difference, 36 U/mL (36 kU/L; 95% confidence interval [CI], 87 to 159 U/mL [87 to 159 kU/L]). B, Mean difference, 11 U/mL (11 kU/L; 95% CI, 98 to 76 U/mL [98 to 76 kU/L]). C, Mean difference, 63 U/mL (63 kU/L; 95% CI, 69 to 194 U/mL [69 to 194 kU/L]). D, Mean difference, 24 U/mL (24 kU/L; 95% CI, 96 to 143 U/mL [96 to 143 kU/L]). E, Mean difference, 6 U/mL (6 kU/L; 95% CI, 94 to 105 U/mL [94 to 105 kU/L]). F, Mean difference, 30 U/mL (30 kU/L; 95% CI, 158 to 99 U/mL [158 to 99 kU/L]). Access 2, Beckman Coulter, Brea, CA; ADVIA Centaur, Bayer Diagnostics, Tarrytown, NY; ARCHITECT i 2000, Abbott Diagnostics, Abbott Park, IL; AxSYM, Abbott Diagnostics; Elecsys 2010, Roche Diagnostics, Indianapolis, IN; IMMULITE 2000, Diagnostic Products, Los Angeles, CA; VITROS ECi, Ortho Clinical Diagnostics, Raritan, NJ.

The results of the present study show that all 7 secondgeneration immunoassays are highly comparable to each other. The slope of IMMULITE 2000 compared with the VITROS ECi was 0.87. The lower slope of the IMMULITE 2000 method relative to the comparison method could be due partly to a deviation from linearity with underrecovery at CA 125 concentrations between 400 and 500 U/mL (400-500 kU/L). However, results from reference interval studies also gave lower values with the IMMULITE 2000 assay, suggesting differences
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in calibration. The ARCHITECT i2000 had a slope of 1.19 relative to the VITROS ECi comparison method. Results from reference interval studies demonstrated higher values as well. Relatively small differences in calibration likely explain these regression and difference plot results. A previous report that examined 7 CA 125 immunoassays found slopes from regression analysis that ranged between 0.61 and 1.80.13 Our results suggest that agreement among commercial methods has improved since that study was published.
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Table 2 Analytic Concordance of Six Automated CA 125 Immunoassays With the VITROS ECi Comparison Method*
VITROS ECi <35 U/mL Access 2 <35 U/mL 35 U/mL Total ADVIA Centaur <30.2 U/mL 30.2 U/mL Total ARCHITECT i 2000 <35 U/mL 35 U/mL Total AxSYM <35 U/mL 35 U/mL Total Elecsys 2010 <35 U/mL 35 U/mL Total IMMULITE 2000 <21 U/mL 21 U/mL Total
*

35 U/mL 0 79 79 0 79 79 0 79 79 0 79 79 0 79 79 0 79 79

Total

16 3 19 16 3 19 15 4 19 18 1 19 14 5 19 15 4 19

16 82 98 16 82 98 15 83 98 18 80 98 14 84 98 15 83 98

Overall concordance with VITROS ECi was as follows: Access 2, 95/98 (96.9%); ADVIA Centaur, 95/98 (96.9%); ARCHITECT i2000, 94/98 (95.9%); AxSYM, 97/98 (99.0%); Elecsys 2010, 93/98 (94.9%); and IMMULITE 2000, 94/98 (95.9%). Values are given in conventional units; to convert to Systme International units (kU/L), multiply by 1. Access 2, Beckman Coulter, Brea, CA; ADVIA Centaur, Bayer Diagnostics, Tarrytown, NY; ARCHITECT i2000, Abbott Diagnostics, Abbott Park, IL; AxSYM, Abbott Diagnostics; Elecsys 2010, Roche Diagnostics, Indianapolis, IN; IMMULITE 2000, Diagnostic Products, Los Angeles, CA; VITROS ECi, Ortho Clinical Diagnostics, Raritan, NJ.

The large differences observed between methods for some individual patient samples, which are best seen in Figure 2, could be because of the use of different antibodies or unique components of these patient samples. Therefore, even though on average the results agree fairly well across assays, when changing assays for clinical use, we recommend parallel testing by the old and new methods to establish a new baseline for each patient.

Although it is not recommended to use CA 125 for ovarian cancer population screening owing to a lack of adequate sensitivity and specificity for a disease with a relatively low prevalence, it is interesting to examine the effects of assay differences on reference intervals. The 97.5 percentile upper reference limits that were determined using the same group of 120 apparently healthy female subjects ranged from 22 U/mL (22 kU/L) for IMMULITE 2000 to 45 U/mL (45 kU/L) for the ARCHITECT i2000 method. These differences are larger than would be expected based on the method comparison studies discussed previously. The percentage of results obtained for these subjects that were less than 35 U/mL (<35 kU/L) ranged from 94% for the ARCHITECT i2000 to 99% for the IMMULITE 2000. These results suggest that there could be intermethod differences in clinical sensitivity and specificity. The intermethod analytic concordance for 98 samples used in the method comparison study was consistent with our reference interval studies. However, the manufacturers recommend cutoff values of 21 U/mL (21 kU/L) and 30.2 U/mL (30.2 kU/L) for the IMMULITE 2000 and ADVIA Centaur methods, respectively. Comparing the analytic concordance using the manufacturer-recommended cutoff gave overall concordances with the VITROS ECi comparison method ranging from 94.9% (Elecsys 2010) to 99.0% (AxSYM). All methods for CA 125 generally show good performance characteristics and compare reasonably well with each other, especially considering the lack of an internationally recognized standard reference material. However, for some patient samples, differences are observed between methods. These may be because of the use of different antibodies or other factors unique to the reagents of each method or individual sample. Methods should not be used interchangeably, but rather patients should have a new baseline established by parallel testing with the old and new methods.
From the 1Department of Pathology, University of Utah Health Sciences Center, and 2ARUP Institute for Clinical and Experimental Pathology, Salt Lake City.

Table 3 Summary of Information From Reference Population*

Method Access 2 ADVIA Centaur ARCHITECT i 2000 AxSYM Elecsys 2010 IMMULITE 2000 VITROS ECi
* Values

Median CA 125 Concentration (U/mL) 12.0 11.4 16.1 11.8 17 .2 6.3 8.9

97.5 Percentile Upper Reference Limit (U/mL) 37 32 45 34 40 22 29

Reference Samples With CA 125 Concentrations 35 U/mL (%) 97 97 94 97 95 99 97

Range of CA 125 Concentrations (U/mL) 3-80 3-69 4-91 3-62 2-83 1-45 2-60

are given in conventional units; to convert to Systme International units (kU/L), multiply by 1. Access 2, Beckman Coulter, Brea, CA; ADVIA Centaur, Bayer Diagnostics, Tarrytown, NY; ARCHITECT i2000, Abbott Diagnostics, Abbott Park, IL; AxSYM, Abbott Diagnostics; Elecsys 2010, Roche Diagnostics, Indianapolis, IN; IMMULITE 2000, Diagnostic Products, Los Angeles, CA; VITROS ECi, Ortho Clinical Diagnostics, Raritan, NJ.

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Supported by Abbott Diagnostics and the ARUP Institute for Clinical & Experimental Pathology. Address reprint requests to Dr Roberts: c/o ARUP Laboratories, 500 Chipeta Way, Salt Lake City, UT 84108.

References
1. Bast RC Jr, Feeney M, Laarus H, et al. Reactivity of a monoclonal antibody with human ovarian carcinoma. J Clin Invest. 1981;68:1331-1337. 2. Nustad K, Bast RC Jr, OBrien TJ, et al. Specificity and affinity of 26 monoclonal antibodies against the CA 125 antigen: first report from the ISOBM TD-1 workshop. Tumour Biol. 1996;17:196-219. 3. Bischof P. What do we know about the origin of CA 125? Eur J Obstet Gynecol Reprod Biol. 1993;49:93-98. 4. Kabawat SE, Bast RC Jr, Bhan AK, et al. Tissue distribution of a coelomic-epitheliumrelated antigen recognized by the monoclonal antibody OC125. Int J Gynecol Pathol. 1983;2:275-285. 5. Chan DW, Booth RA, Diamandis EP. Tumor markers. In: Burtis CA, Ashwood ER, Bruns DE, eds. Tietz Textbook of Clinical Chemistry and Molecular Diagnostics. 4th ed. Philadelphia, PA: Saunders; 2005:745-795. 6. Carlson KJ, Skates SJ, Singer DE. Screening for ovarian cancer. Ann Intern Med. 1994;121:124-132.

7. Van Kamp GJ, Verstraeten AA, Kenemans P. Discordant serum CA 125 values in commercial immunoassays. Eur J Obstet Gynecol Reprod Biol. 1993;49:99-103. 8. Kenemans P, Bon GG, Kessler AC, et al. Multicenter technical and clinical evaluation of a fully automated enzyme immunoassay for CA 125. Clin Chem. 1992;38:1466-1471. 9. Kenemans P, van Kemp GJ, Oehr P, et al. Heterologous double determinant immunoradiometric assay CA 125, II: reliable second-generation immunoassay for determining CA 125 in serum. Clin Chem. 1993;39:2509-2513. 10. Uhl W, Denk B. Improved CA 125 determinations using two different monoclonal antibodies. In: Klapdor R, ed. Current Tumor Diagnosis: Applications, Clinical Relevance, Research Trends. Munich, Germany: Zuckschwerdt Verlag; 1994:384388. 11. Kenemans P, Verstraeten AA, van Kamp GJ, et al. The second generation CA 125 assays. Ann Med. 1995;27:107-113. 12. Bonfrer JM, Baan AW, Jansen E, et al. Technical evaluation of three second generation CA 125 assays. Eur J Clin Chem Clin Biochem. 1994;32:201-207. 13. Davelaar EM, van Kamp GJ, Verstraeten AA, et al. Comparison of seven immunoassays for quantification of CA 125 in serum. Clin Chem. 1998;44:1417-1412.

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