Maillet M, Maubon D, Brion J P, François P, Molina L, Stahl J P, Epaulard O, Bosseray A, Pavese P
Infectious Diseases Department, CHU de Grenoble, BP 218, 38 043, Grenoble cedex 9, France,
Eur J Clin Microbiol Infect Dis. 2014 Mar;33(3):331-6. doi: 10.1007/s10096-013-1960-3. Epub 2013 Aug 30.
Conventional polymerase chain reaction (PCR) in respiratory samples does not differentiate between Pneumocystis pneumonia (PCP) and Pneumocystis jirovecii (Pj) colonization. We used Pj real-time quantitative PCR (qPCR) with the objective to discriminate PCP from Pj colonization in immunocompromised patients. All positive Pj qPCR [targeting the major surface glycoprotein (MSG) gene] obtained in respiratory samples from immunocompromised patients presenting pneumonia at the Grenoble University Hospital, France, were collected between August 2009 and April 2011. Diagnoses were retrospectively determined by a multidisciplinary group of experts blinded to the Pj qPCR results. Thirty-one bronchoalveolar lavages and four broncho aspirations positive for the Pj qPCR were obtained from 35 immunocompromised patients. Diagnoses of definite, probable, and possible PCP, and pneumonia from another etiology were retrospectively made for 7, 4, 5, and 19 patients, respectively. Copy numbers were significantly higher in the "definite group" (median 465,000 copies/ml) than in the "probable group" (median 38,600 copies/ml), the "possible group" (median 1,032 copies/ml), and the "other diagnosis group" (median 390 copies/ml). With the value of 3,160 copies/ml, the sensitivity and specificity of qPCR for the diagnosis of PCP were 100 % and 70 %, respectively. With the value of 31,600 copies/ml, the sensitivity and specificity were 80 % and 100 %, respectively. The positive predictive value was 100 % for results with more than 31,600 copies/ml and the negative predictive value was 100 % for results with fewer than 3,160 copies/ml. qPCR targeting the MSG gene can be helpful to discriminate PCP from Pj colonization in immunocompromised patients, using two cut-off values, with a gray zone between them.
呼吸道样本中的传统聚合酶链反应(PCR)无法区分肺孢子菌肺炎(PCP)和耶氏肺孢子菌(Pj)定植。我们采用Pj实时定量PCR(qPCR)来鉴别免疫功能低下患者的PCP和Pj定植情况。收集了2009年8月至2011年4月期间在法国格勒诺布尔大学医院就诊的肺炎免疫功能低下患者呼吸道样本中所有Pj qPCR阳性结果(针对主要表面糖蛋白(MSG)基因)。诊断由对Pj qPCR结果不知情的多学科专家小组进行回顾性判定。从35名免疫功能低下患者中获取了31份支气管肺泡灌洗样本和4份支气管吸取样本,其Pj qPCR结果呈阳性。分别对7、4、5和19名患者进行回顾性诊断,确诊为确诊、可能、疑似PCP以及其他病因引起的肺炎。“确诊组”的拷贝数(中位数为465,000拷贝/毫升)显著高于“可能组”(中位数为38,600拷贝/毫升)、“疑似组”(中位数为1,032拷贝/毫升)和“其他诊断组”(中位数为390拷贝/毫升)。以3,160拷贝/毫升为阈值,qPCR诊断PCP的敏感性和特异性分别为100%和70%。以31,600拷贝/毫升为阈值,敏感性和特异性分别为80%和100%。拷贝数超过31,600拷贝/毫升时阳性预测值为100%,低于3,160拷贝/毫升时阴性预测值为100%。针对MSG基因的qPCR有助于鉴别免疫功能低下患者的PCP和Pj定植情况,使用两个临界值,两者之间存在一个灰色区域。