Abstract
To investigate the potential association between prostate infection and chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS), we used molecular approaches described in previous reports. These methods employed standard polymerase chain (PCR) reaction assays to provide a qualitative evaluation of prostatic bacterial species. Here, we report on the detection of prostatic bacteria using a real-time PCR. Template DNAs were examined from prostatic tissue samples from patients with CP/CPPS. Two PCR primer sets were used: one that amplifies a portion of all known bacterial ribosomal DNAs (16S rDNAs) and one that is specific for Escherichia coli as opposed to related, E. coli-like bacteria. The 16S rDNA real-time PCR assay detected bacterial DNAs in eight (26%) of 31 samples from patients with CP/CPPS, including three samples (10%) that were also positive by the E. coli real-time PCR assay. These E. coli positives were quantified at approximately 103 cfu/ml of tissue digested. Quantification, speed and specificity make real-time PCR a promising approach for the quantitative detection and identification of prostatic bacteria from CP/CPPS patients.
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References
Aldea C, Alvarez CP, Folgueira L, Delgado R, Otero JR (2002) Rapid detection of herpes simplex virus DNA in genital ulcers by real-time PCR using SYBR Green I dye as the detection signal. J Clin Microbiol 40:1060–1062
Bell CA, Uhl JR, Hadfield TL, David JC, Meyer RF, Smith TF, Cockerill III FR (2002) Detection of Bacillus anthracis DNA by LightCycler PCR. J Clin Microbiol 40:2897–2902
Bialek R, Weiss M, Bekure-Nemariam K, Najvar LK, Alberdi MB, Graybill JR, Reischl U (2002) Detection of Cryptococcus neoformans DNA in tissue samples by nested and real-time PCR assays. Clin Diagn Lab Immunol 9:461–469
Carattoli A, Dionisi AM, Luzzi I (2002) Use of a LightCycler gyrA mutation assay for identification of ciprofloxacin-resistant Campylobacter coli. FEMS Microbiol Lett 214:87–93
Collins MM, Stafford RS, O'Leary MP, Barry MJ (1998) How common is prostatitis? A national survey of physician visits. J Urol 159:1224-1228
He Q, Wang JP, Osato M, Lachman LB (2002) Real-time quantitative PCR for detection of Helicobacter pylori. J Clin Microbiol 40:3720–3728
Hochreiter WW, Duncan JL, Schaeffer AJ (2000) Evaluation of the bacterial flora of the prostate using a 16S rRNA gene based polymerase chain reaction. J Urol 163:127–130
Keay S, Zhang CO, Baldwin BR, Alexander RB (1999) Polymerase chain reaction amplification of bacterial 16S rRNA genes in prostate biopsies from men without chronic prostatitis. Urology 53:487–491
Krieger JN, Eagan K (1991) Comprehensive evaluation and treatment of 75 men referred to chronic prostatitis clinic. Urology 38:11–19
Krieger JN, Riley DE, Roberts MC, Berger RE (1996) Prokaryotic DNA sequences in patients with chronic idiopathic prostatitis. J Clin Microbiol 34:3120–3128
Krieger JN, Nyberg LJr, Nickel JC (1999) NIH consensus definition and classification of prostatitis. JAMA 282:236–237
Krieger JN, Riley DE, Vesella RL, Miner D, Ross SO, Lange PH (2000) Bacterial DNA sequences in prostate tissue from patients with prostate cancer and chronic prostatitis. J Urol 164:1221–1228
Naber KG, Weidner W (2000) Chronic prostatitis—an infectious disease. J Antimicrob Chemother 46:157–161
Nickel JC, Downey J, Johnston B, Clark J, The Canadian Prostatitis Research Group (2001) Predictors of patient response to antibiotic therapy for the chronic prostatitis/chronic pelvic pain syndrome: a prospective multicenter clinical trial. J Urol 165:1539–1544
Palladino S, Kay I, Fonte R, Flexman J (2001) Use of real-time PCR and the LightCycler system for the rapid detection of Pneumocystis carinii in respiratory specimens. Diagn Microbiol Infect Dis 39:233–236
Rantakokko-Jalava K, Jalava J (2001) Development of conventional and real-time PCR assays for detection of Legionella DNA in respiratory specimens. J Clin Microbiol 39:2904–2910
Reischl U, Linde HJ, Lehn N, Landt O, Barratt K, Wellinghausen N (2002) Direct detection and differentiation of Legionella spp. and Legionella pneumophila in clinical specimens by dual-color real-time PCR and melting curve analysis. J Clin Microbiol 40:3814–3817
Riley DE, Berger RE, Miner DC, Krieger JN (1998) Diverse and related 16S rRNA-encoding DNA sequences in prostate in prostate tissues of men with chronic prostatitis. J Clin Microbiol 36:1646–1652
Stamey TA, Fair WR, Timothy MM, Chung HK (1968) Antibacterial nature of prostatic fluid. Nature 218:444–447
Whiley DM, LeCornec GM, Mackay IM, Siebert DJ, Sloots TP (2002) A real-time PCR assay for the detection of Neisseria gonorrhoeae by LightCycler. Diagn Microbiol Infect Dis 42:85–89
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This study was supported in part by grant DK38955 from the National Institutes of Health, Bethesda, MD, and a Veteran Affairs Merit Review Award, Bacterial DNA in Prostate Disease.
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Takahashi, S., Riley, D.E. & Krieger, J.N. Application of real-time polymerase chain reaction technology to detect prostatic bacteria in patients with chronic prostatitis/chronic pelvic pain syndrome. World J Urol 21, 100–104 (2003). https://doi.org/10.1007/s00345-003-0326-3
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DOI: https://doi.org/10.1007/s00345-003-0326-3