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对于未感染HIV的免疫功能低下患者,实时荧光定量聚合酶链反应(Real-time PCR)在肺孢子菌肺炎诱导痰诊断方面比传统聚合酶链反应更具特异性。

Real-time PCR is more specific than conventional PCR for induced sputum diagnosis of Pneumocystis pneumonia in immunocompromised patients without HIV infection.

作者信息

Fujisawa Tomoyuki, Suda Takafumi, Matsuda Hiroyuki, Inui Naoki, Nakamura Yutaro, Sato Jun, Toyoshima Mikio, Nakano Yutaka, Yasuda Kazumasa, Gemma Hitoshi, Hayakawa Hiroshi, Chida Kingo

机构信息

Second Division, Department of Internal Medicine, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka, Japan.

出版信息

Respirology. 2009 Mar;14(2):203-9. doi: 10.1111/j.1440-1843.2008.01457.x. Epub 2008 Dec 11.

Abstract

BACKGROUND AND OBJECTIVE

The diagnosis of Pneumocystis pneumonia (PCP) is based on microscopic examination of respiratory specimens. PCP patients without AIDS have a lower burden of P. jiroveci than those with AIDS, which leads to difficulty in detecting the organisms. Although conventional PCR (c-PCR) has been used to detect the DNA, it is frequently positive in patients with colonization. Real-time PCR (r-PCR), a method to detect the DNA quantitatively, might be helpful in distinguishing between infection and colonization. We investigated the utility of real-time PCR in the diagnosis of PCP in non-AIDS patients.

METHODS

Induced sputum samples obtained from 86 non-HIV immunocompromized patients with clinical symptoms of pulmonary infection were evaluated for the presence of Pneumocystis jiroveci-specific DNA using c-PCR and r-PCR. The diagnosis of PCP was confirmed by typical clinical and radiological findings and response to treatment.

RESULTS

Of the 86 patients, 17 were diagnosed as having PCP. Twenty-eight samples were positive for c-PCR, but the false-positive rate was high (46.4%). Sensitivity, specificity and positive predictive values (PPV) of c-PCR were 88.2%, 81.2% and 53.6%, respectively. Concentrations of the DNA detected by r-PCR were significantly higher in PCP patients than in non-PCP patients. Using 30 copies per tube as a cut-off value for the diagnosis of PCP, the sensitivity (82.4%) of r-PCR was almost equal to c-PCR. Notably, its specificity and PPV were higher than c-PCR (98.6% and 93.3%, respectively).

CONCLUSIONS

r-PCR on induced sputum is more useful for diagnosing PCP than c-PCR in non-HIV immunocompromized patients, especially in terms of distinguishing between colonization and infection.

摘要

背景与目的

肺孢子菌肺炎(PCP)的诊断基于呼吸道标本的显微镜检查。非艾滋病的PCP患者的耶氏肺孢子菌负荷低于艾滋病患者,这导致检测该病原体存在困难。尽管传统PCR(c-PCR)已用于检测DNA,但在定植患者中其结果常为阳性。实时PCR(r-PCR)是一种定量检测DNA的方法,可能有助于区分感染和定植。我们研究了实时PCR在非艾滋病患者PCP诊断中的效用。

方法

对86例有肺部感染临床症状的非HIV免疫受损患者获取的诱导痰标本,使用c-PCR和r-PCR评估耶氏肺孢子菌特异性DNA的存在情况。PCP的诊断通过典型的临床和影像学表现以及对治疗的反应来确认。

结果

86例患者中,17例被诊断为PCP。28份标本c-PCR呈阳性,但假阳性率较高(46.4%)。c-PCR的敏感性、特异性和阳性预测值(PPV)分别为88.2%、81.2%和53.6%。PCP患者中r-PCR检测到的DNA浓度显著高于非PCP患者。以每管30拷贝作为PCP诊断的临界值,r-PCR的敏感性(82.4%)几乎与c-PCR相等。值得注意的是,其特异性和PPV高于c-PCR(分别为98.6%和93.3%)。

结论

对于非HIV免疫受损患者,诱导痰的r-PCR在诊断PCP方面比c-PCR更有用,特别是在区分定植和感染方面。

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