Department of Infectious Diseases, Karolinska University Hospital, Stockholm, Sweden.
J Clin Microbiol. 2014 Jan;52(1):83-9. doi: 10.1128/JCM.01742-13. Epub 2013 Oct 23.
We aimed to compare sputum and nasopharyngeal aspirate (NpA) samples and the PCR gene targets lytA and Spn9802 in quantitative PCR (qPCR) assays for rapid detection of pneumococcal etiology in community-acquired pneumonia (CAP). Seventy-eight adult patients hospitalized for radiologically confirmed CAP had both good-quality sputum and NpA specimens collected at admission. These samples were subjected to lytA qPCR and Spn9802 qPCR assays with analytical times of <3 h. Thirty-two patients had CAP with a pneumococcal etiology, according to conventional diagnostic criteria. The following qPCR positivity rates were noted in CAP cases with and without pneumococcal etiology: 96% and 15% (sputum lytA assay), 96% and 17% (sputum Spn9802 assay), 81% and 11% (NpA lytA assay), and 81% and 20% (NpA Spn9802 assay), respectively. The mean lytA and Spn9802 DNA levels were significantly higher in qPCR-positive sputum samples from cases with pneumococcal etiology than in qPCR-positive sputum samples from CAP cases without pneumococcal etiology or qPCR-positive NpA samples from cases with pneumococcal etiology (P < 0.02 for all comparisons). For detection of pneumococcal etiology, receiver operating characteristic curve analysis showed that sputum specimens were superior to NpA specimens as the sample type (P < 0.02 for both gene targets) and lytA tended to be superior to Spn9802 as the gene target. The best-performing test, the sputum lytA qPCR assay, showed high sensitivity (94%) and specificity (96%) with a cutoff value of 10(5) DNA copies/ml. In CAP patients with good sputum production, this test has great potential to be used for the rapid detection of pneumococcal etiology and to target penicillin therapy.
我们旨在比较痰和鼻咽抽吸物(NpA)样本以及聚合酶链反应(PCR)基因靶标 lytA 和 Spn9802 在定量 PCR(qPCR)检测中的作用,以快速检测社区获得性肺炎(CAP)中的肺炎球菌病因。78 名因影像学证实的 CAP 住院的成年患者在入院时采集了高质量的痰和 NpA 标本。这些样本进行了 lytA qPCR 和 Spn9802 qPCR 检测,分析时间<3 小时。根据传统诊断标准,32 名患者患有 CAP 并有肺炎球菌病因。在有和没有肺炎球菌病因的 CAP 病例中观察到以下 qPCR 阳性率:96%和 15%(痰 lytA 检测)、96%和 17%(痰 Spn9802 检测)、81%和 11%(NpA lytA 检测)和 81%和 20%(NpA Spn9802 检测)。具有肺炎球菌病因的 CAP 病例中 qPCR 阳性的痰样本的 lytA 和 Spn9802 DNA 水平明显高于无肺炎球菌病因的 CAP 病例中 qPCR 阳性的痰样本或具有肺炎球菌病因的 qPCR 阳性的 NpA 样本(所有比较均 P<0.02)。对于肺炎球菌病因的检测,受试者工作特征曲线分析表明,与 NpA 样本相比,痰样本作为样本类型具有优势(两个基因靶标均 P<0.02),并且 lytA 作为基因靶标优于 Spn9802。表现最佳的测试是痰 lytA qPCR 检测,其在 10(5)DNA 拷贝/ml 的截断值下具有高灵敏度(94%)和特异性(96%)。在有良好痰产生的 CAP 患者中,该测试具有很大的潜力用于快速检测肺炎球菌病因并针对青霉素治疗。