| ObjectiveTo evaluation the clinical significance of FISH technique for screening bladder transitional cell carcinoma with hematuria.To provide a new method for early diagnosis of bladder cancer with cytology of the urine.Methods(1) We selected 20 relatively healthy people as control group. The special urine examination method detects genetic alterations of the urothelial cells found in the urine of the control group, using fluorescent direct labeled DNA probes binding to the peri-centromeric regions of chromosomes 3, 7 and 17 as well as on the 9p21 locus, respectively. Thresholds were then established.(2) 100 early morning urine samples collecting for 100 patients with hematuria underwent cytology of the urine and FISH examination, respectively. The sensitivity and specificity of both were analyzed statistically. We aimed to evaluate the utility of FISH technique for diagnosing bladder cancer.(3) To explore the applicable conditions for domestic FISH probes and review the advantage and disadvantage for further application. Then try to establish the technological platforms for FISH used in the urinary cytology with standard criteria.Results(1)We statistically analyzed the proportion of cells with abnormal signal in the control group and determined the thresholds. There are at least 100 cells in one slice. The thresholds famula: the thresholds=average value+3*standard deviation. The complete absent, the partial absent and amplification thresholds for P16 gene detected by probes were 3.9%, 5.4% and 3.2% respectively. The absent and amplification thresholds for chromosome 3 were 4.0% and 3.5%; 4.0 % and 2.8% for chromosome 7; 5.8% and 3.4% for chromosome 17 respectively. At least 2 results detected by the probes were greater than the thresholds or there were at least 2 kinds of abnormalities as positive result detected by the probes.(2) For these 100 patients with hematuria, there were 78 patients diagnosed with bladder cancer; 2 were suspected, otherwise were normal, comferme d by pathology results. 44 cases of bladder transitional cell carcinoma were detected with FISH positive in 47 cases, while 22 cases by urinary cytology examination. 8 cases were detected with FISH positive among the 20 cases with Papillary Urothelial Neoplasm of Low Malignant Potential, while 4 of 7 patients with inverted papilloma and 1 patient of suspected bladder cancer. For patients with non transitional cell carcinoma, there was no positive result shown by urinary cytology. The study rated the specificity and sensitivity of the technique 93.6% and75.5%, respectively and sequently 6.4%, 24.5% for false negative and false positive, respectively. The study rated the specificity and sensitivity of the urinary exfoliative cytology 46.8%% and 100%, respectively.(3) There are something more should be done to improve FISH probes. For example, It is of importance to deal with the foreign substance while deal with the cells. More collagenase B or time prolonged is needed. The amount of pepsin and the interacting time also should be carefully controlled. For the temperature, we appreciate 3~5°elevated.Conclusion(1) This new non-invasive diagnostic application of the FISH technique with high detection rate in the field of urology is prior to urinary cytology.(2) FISH technique is of high sensitivity but lower specificity than urinary cytology which may be related to the thresholds. Nevertheless, FISH technique can be performed first, compared to the cystoscopy.(3) More clinical researches should be done to evaluate whether the FISH tequnique can replace the cystoscopy because of the complex and lack of criteria. For the domestic FISH probes, more researches are needed to improve the stability and the reliability, making a possibility for future application.
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