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使用基于16S rRNA基因的聚合酶链反应评估前列腺的细菌菌群。

Evaluation of the bacterial flora of the prostate using a 16S rRNA gene based polymerase chain reaction.

作者信息

Hochreiter W W, Duncan J L, Schaeffer A J

机构信息

Department of Urology, Northwestern University Medical School, Chicago, Illinois, USA.

出版信息

J Urol. 2000 Jan;163(1):127-30.

Abstract

PURPOSE

The role of bacteria in the chronic pelvic pain syndrome (nonbacterial prostatitis and prostatodynia) is controversial and difficult to assess because the bacterial flora of the prostate is not well defined. Polymerase chain reaction (PCR) is a highly sensitive molecular method of bacterial detection. It confirms the sterility of tissue with a high level of confidence and detects small numbers of microbial agents that may represent pathogens. We performed PCR to determine bacterial colonization of the prostate in presumably healthy men and in those undergoing simple or radical prostatectomy.

MATERIALS AND METHODS

We analyzed 28 prostate samples from 18 organ donors from whom prostate tissue was obtained under sterile surgical conditions at organ withdrawal, 14 sterile surgical prostate specimens from 7 patients undergoing radical prostatectomy for prostate cancer who previously underwent transrectal biopsy and 6 sterile surgical specimens from 2 men who underwent simple prostatectomy for benign prostatic hyperplasia (BPH), including 1 with an indwelling catheter for several weeks. For PCR we used 2 sets of primers to detect bacterial 16S rRNA gene sequences. Normal prostate tissue seeded in vitro with known numbers of Escherichia coli was used to assess the sensitivity of PCR.

RESULTS

Only 3 of the 28 organ donor samples had histological signs of minimal inflammation and all other samples appeared to be normal without evidence of inflammatory reaction. All of these samples were PCR negative. Of several PCR control reactions the mixture of prostate tissue seeded with known numbers of E. coli demonstrated the high sensitivity of the assay, allowing the detection of as few as 6 bacteria in the presence of 25 mg. of prostate tissue. A focal and heterogeneous distribution of inflammation and infection was noted in the 14 radical prostatectomy specimens. In the prostate cancer and BPH groups there was a strong association of inflammation with positive PCR findings. Of 11 samples 3 without but all 9 with inflammation were PCR positive.

CONCLUSIONS

PCR is a highly sensitive method for detecting bacteria in the prostate. In our study negative PCR reactions in the prostate tissue of apparently healthy men made the presence of normal bacterial flora in the prostate extremely unlikely. The presence of bacteria and/or inflammation in radical prostatectomy specimens was found to be a localized process. Concordance between inflammation and positive PCR results in simple and radical prostatectomy specimens suggests that bacteria may frequently have a role in histologically inflammatory prostatitis.

摘要

目的

细菌在慢性盆腔疼痛综合征(非细菌性前列腺炎和前列腺痛)中的作用存在争议且难以评估,因为前列腺的细菌菌群尚未明确界定。聚合酶链反应(PCR)是一种高度敏感的细菌检测分子方法。它能高度可靠地确认组织无菌,并检测出可能代表病原体的少量微生物。我们进行PCR以确定健康男性以及接受单纯前列腺切除术或根治性前列腺切除术男性的前列腺细菌定植情况。

材料与方法

我们分析了18例器官捐献者的28份前列腺样本,这些样本是在器官捐献时在无菌手术条件下获取的;7例因前列腺癌接受根治性前列腺切除术且之前接受过经直肠活检的患者的14份无菌手术前列腺标本;以及2例因良性前列腺增生(BPH)接受单纯前列腺切除术的男性的6份无菌手术标本,其中1例留置导尿管数周。对于PCR,我们使用2组引物来检测细菌16S rRNA基因序列。用已知数量的大肠杆菌体外接种正常前列腺组织以评估PCR的敏感性。

结果

28份器官捐献者样本中只有3份有轻微炎症的组织学迹象,其他所有样本看起来正常,无炎症反应迹象。所有这些样本PCR均为阴性。在几个PCR对照反应中,接种已知数量大肠杆菌的前列腺组织混合物显示出该检测方法的高敏感性,在存在25毫克前列腺组织的情况下能检测到低至6个细菌。在14份根治性前列腺切除术标本中发现炎症和感染呈局灶性和异质性分布。在前列腺癌和BPH组中,炎症与PCR阳性结果密切相关。11份样本中,3份无炎症的样本PCR阴性,而9份有炎症的样本PCR均为阳性。

结论

PCR是检测前列腺中细菌的高度敏感方法。在我们的研究中,明显健康男性前列腺组织的PCR阴性反应表明前列腺中存在正常细菌菌群的可能性极小。在根治性前列腺切除术标本中发现细菌和/或炎症是一个局部过程。单纯前列腺切除术和根治性前列腺切除术标本中炎症与PCR阳性结果的一致性表明,细菌可能在组织学上有炎症的前列腺炎中经常起作用。

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