Diagnostic use of PCR for detection of Pneumocystis carinii in oral wash samples
Diagnostic use of PCR for detection of Pneumocystis carinii in oral wash samples
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DOI：
10.1128/jcm.36.7.2068-2072.1998
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发表时间：
1998-07-01
影响因子：
9.4
通讯作者：
Lundgren, B
中科院分区：
文献类型：
--
作者：
Helweg-Larsen, J;Jensen, JS;Lundgren, B
需要开发无创方法来诊断无法进行支气管镜检查或诱导痰的患者的卡氏肺孢子虫肺炎，口腔冲洗标本易于获得，并且可以通过 PCR 扩增和证明卡氏肺孢子虫核酸。在常规临床使用中，需要简单的样本处理和单轮 PCR 来确保快速分析并降低污染风险，我们开发了单轮 Touchdown PCR (TD-PCR) 方案，能够检测样本中的 PCR 抑制。 TD-PCR 在常规诊断实验室进行评估，并与之前描述的在研究实验室运行的 PCR 方案（线粒体 RNA）进行比较。两种 PCR 方法均扩增了 P. carinii 的线粒体 rRNA 基因序列，包括来自 76 名接受诊断性支气管镜检查的连续人类免疫缺陷病毒 1 型感染者的配对支气管肺泡灌洗 (BAL) 和口腔洗液样本。 TD-PCR 程序比线粒体 PCR 程序更快（
There is a need to develop noninvasive methods for the diagnosis of Pneumocystis carinii pneumonia in patients unable to undergo bronchoscopy or induction sputum, Oral wash specimens are easily obtained, and P. carinii nucleic acid can be amplified and demonstrated by PCR In routine clinical use, easy sample processing and single-round PCR are needed to ensure rapid analysis and to reduce the risk of contamination, We developed a single-round Touchdown PCR (TD-PCR) protocol with the ability to detect PCR inhibition in the specimen. The TD-PCR was evaluated in a routine diagnostic laboratory and was compared to a previously described PCR protocol (mitochondrial RNA) run in a research laboratory. Both PCR methods amplified a sequence of the mitochondrial rRNA gene of P. carinii, Paired bronchoalveolar Lavage (BAL) and oral wash specimens from 76 consecutive human immunodeficiency virus type 1-infected persons undergoing a diagnostic bronchoscopy were included. The TD-PCR procedure was quicker than the mitochondrial PCR procedure (