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首页> 外文期刊>Scandinavian journal of infectious diseases. >Rapid simple and nested polymerase chain reaction for the diagnosis of Pneumocystis carinii pneumonia.
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Rapid simple and nested polymerase chain reaction for the diagnosis of Pneumocystis carinii pneumonia.

机译：快速简单的巢式聚合酶链反应用于诊断卡氏肺孢子虫肺炎。

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摘要

We have developed a rapid and easy extraction procedure for polymerase chain reaction (PCR) protocols. Using this simplified step, we evaluated the sensitivity and the specificity of a simple PCR using the primers of Wakefield et al, and of a nested PCR, using new internal primers selected by us, in a total of 89 bronochoalveolar lavage (BAL) fluid samples from 43 immunosuppressed patients. In 13 patients, Pneumocystis carinii pneumonia (PCP) was diagnosed by immunofluorescent antibody (IFA) staining performed on BAL cells cytospun on microscope slides. In seven of these patients we attempted to estimate the post-treatment persistence of P. carinii in BAL, by PCR. After a rapid 2-h extraction procedure, simple and nested PCR were positive in all cases of PCP. SImple and nested PCR both had a 100% sensitivity and a 98 and 84% specificity respectively, compared to IFA. After completion of treatment, BAL liquids from asymptomatic patients were no longer positive by both PCR techniques, whereas the BAL fluid of a patient who was still symptomatic was positive by simple and nested PCR. In follow-up BAL fluids of patients with proven PCP, persistence of P. carinii was detected for a longer period by nested PCR than by simple PCR. Simple PCR is a very rapid and sensitive assay for the diagnosis of PCP in BAL fluid and gives clear-cut results in the case of doubtful IFA staining results. Nested PCR seems to improve the sensitivity of the detection of P. carinii in BAL fluid, but the clinical relevance of a positive result remains to be investigated..
机译：我们为聚合酶链反应（PCR）方案开发了一种快速简便的提取程序。使用此简化步骤，我们评估了使用Wakefield等人引物进行的简单PCR以及使用我们选择的新内部引物进行的巢式PCR的敏感性和特异性，共计89个支气管肺泡灌洗液（BAL）来自43名免疫抑制患者。在13例患者中，通过对显微镜载玻片上纺丝的BAL细胞进行免疫荧光抗体（IFA）染色，诊断出卡氏肺孢子虫肺炎（PCP）。在其中的7位患者中，我们尝试通过PCR估计BAL中卡氏疟原虫的治疗后持续性。经过2小时的快速提取程序后，在所有PCP病例中，简单的巢式PCR均为阳性。与IFA相比，简单和巢式PCR分别具有100％的敏感性和98％和84％的特异性。在完成治疗后，两种PCR技术均无症状患者的BAL液不再呈阳性，而仍存在症状的患者的BAL液通过简单的巢式PCR呈阳性。在经过证实的PCP患者的随访BAL液中，巢式PCR检出的Carinii持久性比简单PCR检出的时间更长。简单PCR是用于诊断BAL液中PCP的非常快速和灵敏的检测方法，如果IFA染色结果有疑问，则可以得到清晰的结果。巢式PCR似乎可以提高检测BAL液中卡氏疟原虫的灵敏度，但阳性结果的临床相关性尚待研究。

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