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通过聚合酶链反应评估尿液样本中伯氏疏螺旋体DNA的检测情况。

Evaluation of the detection of Borrelia burgdorferi DNA in urine samples by polymerase chain reaction.

作者信息

Maiwald M, Stockinger C, Hassler D, von Knebel Doeberitz M, Sonntag H G

机构信息

Hygiene Institut der Universität, Heidelberg, Germany.

出版信息

Infection. 1995 May-Jun;23(3):173-9. doi: 10.1007/BF01793860.

Abstract

It is difficult in some cases to identify an infection caused by Borrelia burgdorferi and to monitor the effect of therapy. Seropositivity will persist even after successful treatment and therefore may suggest ongoing infection. For direct detection of B. burgdorferi DNA in human urine samples, the polymerase chain reaction (PCR) was evaluated. A published primer system was selected, which amplifies a 259 bp fragment from the gene encoding the 23S rRNA. The lower detection limit of the primer system was 10 fg of extracted B. burgdorferi DNA. Several methods for the pretreatment of urine samples were tested. Of these, the Geneclean kit (Bio 101, USA) showed the best results. A total of 114 urine samples from 74 patients belonging to three clinical groups was investigated: (i) 51 samples from 26 patients with active Lyme disease, (ii) 36 samples from 27 patients with previous infection but no symptoms at the time the urine was collected, and (iii) 27 samples from 21 seronegative control patients without Lyme disease. B. burgdorferi DNA was detected in 25 urine samples of 17 patients with active disease, whereas 26 samples from this group of patients were negative. Only one asymptomatic case with previous infection showed a positive result, and the urine samples of the patients without Lyme disease were uniformly negative. Two of four patients from whom samples before and directly after onset of therapy were available converted from negative to positive PCR results after initiation of therapy, accompanied by the symptoms of a Jarisch-Herxheimer reaction.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

在某些情况下,很难识别由伯氏疏螺旋体引起的感染并监测治疗效果。即使治疗成功,血清阳性仍会持续存在,因此可能提示存在持续感染。为了直接检测人尿液样本中的伯氏疏螺旋体DNA,对聚合酶链反应(PCR)进行了评估。选择了一种已发表的引物系统,该系统可从编码23S rRNA的基因中扩增出一个259 bp的片段。该引物系统的检测下限为10 fg提取的伯氏疏螺旋体DNA。测试了几种尿液样本预处理方法。其中,Geneclean试剂盒(美国Bio 101公司)显示出最佳结果。共调查了来自三个临床组的74例患者的114份尿液样本:(i)来自26例活动性莱姆病患者的51份样本,(ii)来自27例既往感染但在采集尿液时无症状的患者的36份样本,以及(iii)来自21例无莱姆病的血清阴性对照患者的27份样本。在17例活动性疾病患者的25份尿液样本中检测到伯氏疏螺旋体DNA,而该组患者的26份样本为阴性。只有1例既往感染的无症状病例呈阳性结果,无莱姆病患者的尿液样本均为阴性。在4例治疗开始前和治疗后直接采集样本的患者中,有2例在治疗开始后PCR结果从阴性转为阳性,并伴有雅里希-赫克斯海默反应的症状。(摘要截短至250字)

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