Establishment and application of DD3 gene detection with real-time fluores-cence quantified reverse transcription polymerase chain reaction in patients with prostate cancer
AIM: To establish the method of detecting DD3 gene with real-time fluorescence quantified reverse transcription polymerase chain reaction and to employ it in the early diagnosis of prostate cancer. METHODS: Specific primer and marked probes for DD3 gene were designed according to full DD3 mRNA sequence in Cenbank. The vector to prepare the quantitative standard for DD3 gene detection was constructed and quantitative real-time fluorescence methods were employed to detect blood, urine and prostatic fluid specimens. RESULTS: By laboratory evaluation of the stability, specificity, repeatability and sensitivity, the lowest limit for detection of DD3 gene by real-time fluorescence quantitative PCR was 1.64 × 103 copy/L. Its line scope ranged from 1.64 × 103 to 1.64 × 1015 copy/L. The specimens of blood, urine and prostatic fluid from 19 patients with prostate cancers, 20 non-prostate cancer patients and 30 healthy volunteers were assessed. The positive rate was 100% (blood), 80% (urine)and 100% (prostatic fluid). Among the prostate gland liquid specimens from the 20 non-prostate cancer patients, only one specimen showed low copy(1 × 105 copy/L). Specimens from the 30 healthy volunteers were all negative. In the specimens of blood, urine and prostatic fluid, the DD3 mRNA content of the prostate cancer patient was significantly higher than that of non-prostate cancer patients and healthy volunteers. (P ＜ 0.05). CONCLUSION: DD3 gene can be used as a specific marker for early prostate cancer diagnosis and it can be used for the examination for blood, seminal fluid, urine and prostatic fluid. The examination can be employed for early prostate cancer diagnosis, micro translocation diagnosis, treatment and prognosis.
NETL
NSTL
万方数据
维普
