title: Detection of tumor markers in prostate cancer and comparison of sensitivity between real time and nested PCR.
abstract: The objective of this study is to investigate and compare the sensitivity in conventional PCR, quantitative real time PCR, nested PCR and western blots for detection of prostate cancer tumor markers using prostate cancer (PCa) cells. We performed conventional PCR, quantitative real time PCR, nested PCR, and western blots using 5 kinds of PCa cells. Prostate specific antigen (PSA), prostate specific membrane antigen (PSMA), and androgen receptor (AR) were compared for their detection sensitivity by real time PCR and nested PCR. In real time PCR, there was a significant correlation between cell number and the RNA concentration obtained (R(2)=0.9944) for PSA, PSMA, and AR. We found it possible to detect these markers from a single LNCaP cell in both real time and nested PCR. By comparison, nested PCR reached a linear curve in fewer PCR cycles than real time PCR, suggesting that nested PCR may offer PCR results more quickly than real time PCR. In conclusion, nested PCR may offer tumor maker detection in PCa cells more quickly (with fewer PCR cycles) with the same high sensitivity as real time PCR. Further study is necessary to establish and evaluate the best tool for PCa tumor marker detection.
authors.affiliation: Infection Center, Kobe University Graduate School of Medicine, Kobe, Japan.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Matsuoka
authors.forename: Katsumi
authors.initials: K
authors.lastname: Shigemura
authors.forename: Fukashi
authors.initials: F
authors.lastname: Yamamichi
authors.forename: Masato
authors.initials: M
authors.lastname: Fujisawa
authors.forename: Masato
authors.initials: M
authors.lastname: Kawabata
authors.forename: Toshiro
authors.initials: T
authors.lastname: Shirakawa
mesh_terms: D014408:Biomarkers, Tumor / Q000378:metabolism*; D002452:Cell Count; D045744:Cell Line, Tumor; D020869:Gene Expression Profiling; D006801:Humans; D008297:Male; D011471:Prostatic Neoplasms / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D012680:Sensitivity and Specificity
chemical_list: D014408:Biomarkers, Tumor; D012333:RNA, Messenger