HOME          Company Info          Press Releases          How To Get Test           Request Information          Comments/Suggestions

PATIENT TEST PROCEDURE

1. Bring test materials and patient urine sample to room temperature (17 - 37°C, 63 - 99°F). Gently swirl patient's urine sample.
2. Remove the test device and dropper from foil package. Throw away small desiccant pouch. Place the device on a clean, well-lit, flat surface and label with the patient's identification.
	3. Set timer. Fill the dropper provided with the patient's urine sample and hold it upright above the sample well as shown.
	4. Allow 5 FULL drops (without air bubbles) to fall into the sample well. Start timer.
	5. When timer reaches 5 minutes, read results within 1 minute. Read results as shown under "Interpretation of Results."
Space Space	6. Discard used dropper and test device in a proper biohazard container.
	Read at 5 minutes but NO LATER THAN 6 MINUTES, Test result is not valid if read later than 6 minutes..

INTERPRETATION OF RESULTS

1. Check the procedural Control () window. A pink or reddish-brown line must appear for the test to be valid.

2. Positive Result: Carefully look at Patient (P) window. ANY pink or reddish-brown colored line, NO MATTER HOW FAINT, in the Patient (P) window is a positive result. Neither the intensity nor the color should be compared to that seen in the procedural Control () window.

3. Negative Result: Carefully look at Patient (P) window. No colored line in the Patient (P) window is a negative result.

4. Invalid Test Result: If no line appears in the procedural Control () window, the test is invalid and must be repeated with a new device. The most common reason for an invalid test result is failure to add exactly 5 FULL drops.

QUALITY CONTROL

Good laboratory practices recommend the use of appropriate controls. There are two types of controls for the BTA stat test - the internal procedural control and external controls.
Procedural Control
The procedural Control is found in the Control () zone of the test device. This control assures the operator that (A) sample addition and migration through the device has occurred and that (B) the control goat anti-mouse antibody and the reporter MAb antibody are intact and functional. This control does not ensure that the Patient (P) zone is accurately detecting the presence or absence of bladder tumor associated antigen in the sample.
External Controls
External controls are used to assure the operator that the capture and conjugated antibodies are present and reactive. External controls will not detect an error in performing the patient test procedure. The BTA stat Test Control Kit (Cat. No. 661105) is available separately and contains Positive and Negative Control solutions.
If controls do not perform as expected, do not use the test results. Repeat the test or call Polymedco Technical Service at 800-431-2123.

LIMITATIONS

Results of the BTA stat test should not be interpreted as absolute evidence for the presence or absence of bladder cancer. Any disease which could cause endogenous hCFH to leak into the bladder may cause a positive test result. Positive results have been observed in some patients with renal stones, nephritis, renal cancer (including upper tract TCC), urinary tract infections, cystitis, sexually transmitted diseases and recent trauma to the bladder or urinary tract. The BTA stat test should not be used as a screening test for individuals without biopsy confirmed bladder cancer. The result from the BTA stat test should be used only in conjunction with information available from the clinical evaluation of the patient and other diagnostic procedures.

EXPECTED RESULTS

CLINICAL SENSITIVITY
BTA stat test sensitivity (Table I) was determined using urine samples from 220 patients with biopsy confirmed bladder tumor recurrence. Samples were collected from 5 different geographic locations throughout the United States and stored frozen until tested. Testing of samples for this study was performed at B.D.S., Inc. The average patient age was 68 years, 79% were males, 67% Caucasian, 1% African American, 4% Asian, Hispanic or other, and 27% of unknown ethnicity. Results are presented below by stage and by grade of the tumor.

Table I. BTA stat TEST SENSITIVITY BY STAGE AND GRADE*

*3 patients without stage and 12 without grade determinations.

Table II presents the overall sensitivity in 220 patients with histologically confirmed bladder cancer recurrence (Table I), as well as the specificity in 107 patients who were being monitored for recurrence of bladder cancer and determined by cystoscopy and/or biopsy to have no evidence of disease (NED) at the time of the BTA stat test determination. 

Table II. BTA stat TEST RESULTS FROM PATIENTS WITH A HISTORY OF BLADDER CANCER

Monitoring sensitivity = 67% (60 - 73, 95% confidence interval)
Monitoring specificity = 70% (61 - 79, 95% confidence interval)

Using the data in Table II and a 10%, 20%, and 30% hypothetical prevalence of bladder cancer recurrence, the positive predictive values and negative predictive values of the BTA stat test are presented in Table III. Due to the possibility that bladder cancer may have been present in some of the NED individuals in this study, yet missed by cystoscopy, the true specificity in these patients and the positive and negative predictive values are likely to be higher. 

Table III. HYPOTHETICAL POSITIVE PREDICTIVE VALUES (PPV)
AND NEGATIVE PREDICTIVE VALUES (NPV) OF THE BTA stat TEST

A subset of the patients with histologically confirmed bladder cancer (131) also had voided urine cytology (VUC) performed on the same sample as the BTA stat test (Table IV). The BTA stat test was shown to be more sensitive than VUC in all categories except for Tis (tumor in situ). 

Table IV. BTA stat TEST AND VUC SENSITIVITIES

In a subset of patients (99) with a history of bladder cancer and no evidence of disease the specificity of the BTA stat test was 69% compared to VUC with a specificity of 97%.

CLINICAL SPECIFICITY
BTA stat test specificity (Table V) was determined using urine samples from 555 individuals with no history of bladder cancer. Samples were collected from 5 different geographic locations throughout the United States and stored frozen (-80°C) until tested. Testing of samples for this study was performed at B.D.S., Inc. The average age was 55 years, 52% were females, 86% were Caucasian, 8% African American, 4% Asian, Hispanic or other, and 3% of unknown ethnicity. The normal healthy population consisted of 60% non-smokers. The non-genitourinary (GU) diseases and cancers (71% of samples provided by females) included diabetes, arthritis, lupus erythematosus and other collagen degenerative diseases, as well as leukemia, lymphomas, breast, lung and gastrointestinal cancers. The non-bladder genitourinary cancers category (69% of samples provided by males) consisted of prostate, renal cell, renal TCC, endometrial, ovarian and other GU carcinomas. The GU disease category (52% of samples provided by females) consisted of patients with benign prostatic hyperplasia (BPH), prostatitis, urethritis, renal stones and disease, urinary tract infections (UTI), incontinence, sexually transmitted diseases (STD) and other disorders.
The results indicated that in healthy individuals and individuals without GU diseases and malignancies, the BTA stat test negative rate was 95% and 93%, respectively. Positive BTA stat test results may occur in patients with renal disease such as stones and nephritis and patients with renal cancer including upper tract TCC. Expected results may vary depending on the patient population tested.

Table V. BTA stat TEST SPECIFICITY RESULTS

^A   total of subjects with no history of bladder cancer

^B   No evidence of disease confirmed by cystoscopy and/or biopsy;
78% of patients in this category were males

PERFORMANCE CHARACTERISTICS

HIGH DOSE HOOK EFFECT
High dose hook (prozone) effect tests were conducted to determine if the BTA stat test is free from interference from high concentration positive patient samples. Results showed that there was no prozone effect up to 12,400 U/mL bladder tumor associated antigen in a patient's urine sample, which was the highest concentration available for testing.
REPRODUCIBILITY
Three lots of BTA stat devices were used for the reproducibility studies to determine day-to-day, reader-to-reader and lot-to-lot variability. These studies were conducted by testing 10 replicates of 4 blinded samples per day for 5 days using three independent readers for each lot of devices. Between laboratory reproducibility studies were conducted at three laboratories by testing 10 replicates of 4 blinded samples on one lot of BTA stat devices. All reproducibility studies showed nearly total agreement with the exception of samples near the limit of detection, which is to be expected for qualitative tests.

INTERFERING SUBSTANCES

Normal and TCC positive urine pools containing the substances listed below were tested in the BTA stat test. 

TABLE VI. INTERFERING SUBSTANCES

*    MLT - maximum level tested

^A   Negative Interference: substance decreased the intensity of a TCC positive urine test result

^B   Subjecting samples to one freeze/thaw cycle resulted in no interference at 1.25 x 10¹⁰ CFU/mL, the MLT. 

^C   Results of interference studies unchanged by subjecting samples to one freeze/thaw cycle

^D   Substance’s coloration caused results for both normal and TCC positive urine to be difficult to interpret

REFERENCES

1.  Cancer Facts and Figures, American Cancer Society, 1996.

2.  Thrasher J., Crawford E.: Current Management of Invasive Metastatic Transitional Cell Carcinoma of the Bladder. The Journal of Urology 149:957-972, 1993.

3.  Murphy W.: Current Status of Urinary Cytology in the Evaluation of Bladder Neoplasms. Human Pathology 21:886-896, 1990.

4.  Umiker W.: Accuracy of Cytologic Diagnosis of Cancer of the Urinary Tract. Symposium on Diagnostic Accuracy of Cytologic Technics 8:186-193,1964.

5.  Badalament R.A., Hermansen D.K., Kimmel M., Gay H., Herr H.W., Fair W.R., Whitmore W.F.,Jr., Melamed M.R.: The Sensitivity of Bladder Wash Flow Cytometry, Bladder Wash Cytology, and Voided Cytology in the Detection of Bladder Carcinoma. Cancer 60:1423-1427, 1987.

6.  Sarosdy M.F., Hudson M.A., et al: Improved Detection of Recurrent Bladder Cancer Using the Bard BTA stat Test. Urology 50(3):349 - 353,1997.

7.  Raitanen M.-P., Marttila T., et al: The Bard BTA stat Test in Monitoring of Bladder Cancer. The Journal of Urology 157: 28, 1997.

8.  Kinders R., Jones T., et al: Complement Factor H or a Related Protein Is a Marker for Transitional Cell Carcinoma of the Bladder. Clinical Cancer Research 4:2511-2520, 1998.

9.  Corey M., Kinders R., et al: Factor H Related Proteins Are Upregulated In Bladder Cancer. Proceedings of the American Association for Cancer Research 39: 263, 1998.

10. Austyn J. M., Wood K. J.: Principles of Cellular and Molecular Immunology. Oxford University Press p. 522 -554, 1993.

11. Corey M.J., Kinders R.J., et al: Enhancement of Complement-Mediated Lysis of Cancer Cells By Anti-Factor H Antibodies. Proceedings of the American Association for Cancer Research 39: 304, 1998.