Lipschik G Y, Gill V J, Lundgren J D, Andrawis V A, Nelson N A, Nielsen J O, Ognibene F P, Kovacs J A
Critical Care Medicine Department, Warren G Magnuson Clinical Centre, National Institutes of Health, Bethesda, Maryland 20814.
Lancet. 1992 Jul 25;340(8813):203-6. doi: 10.1016/0140-6736(92)90469-j.
Detection of Pneumocystis carinii by the polymerase chain reaction (PCR) may facilitate non-invasive diagnosis of P carinii pneumonia and study of its epidemiology. We have compared the sensitivity and specificity of two PCR methods with those of conventional staining for detection of P carinii in induced sputum, bronchoalveolar lavage fluid (BAL), and blood. Of 71 sputum samples, 17 were from patients with microbiologically confirmed P carinii pneumonia. A nested PCR method correctly identified the presence of P carinii in all 17 (100% sensitive, 95% confidence interval [CI] 81-100%) and found no organisms in 50 of 54 microbiologically negative samples (93% specific, 95% CI 82-98%). PCR with a single primer pair was 71% sensitive (44-90%) and 94% specific (85-99%). The sensitivity of conventional staining methods (direct and indirect fluorescence antibody and toluidine-blue-O tests) was significantly less (38-53%) than that of nested PCR (p less than 0.05). In BAL, neither PCR method was significantly better than the conventional staining methods. P carinii was detected in BAL or sputum from 10 immunocompromised patients without microbiological evidence of P carinii pneumonia, which suggests that symptom-free carriers or subclinical infection can exist. P carinii was detected by nested PCR in blood from 2 of 3 patients with disseminated pneumocystosis but in only 1 of 11 patients with P carinii infection restricted to the lungs. Nested PCR on induced sputum is more sensitive than conventional staining methods for the diagnosis of P carinii pneumonia and provides a non-invasive method of detecting disseminated disease.
通过聚合酶链反应(PCR)检测卡氏肺孢子虫可能有助于卡氏肺孢子虫肺炎的非侵入性诊断及其流行病学研究。我们比较了两种PCR方法与传统染色法在检测诱导痰、支气管肺泡灌洗(BAL)液和血液中卡氏肺孢子虫的敏感性和特异性。在71份痰标本中,17份来自微生物学确诊为卡氏肺孢子虫肺炎的患者。一种巢式PCR方法正确鉴定出所有17份标本中均存在卡氏肺孢子虫(敏感性100%,95%可信区间[CI]81 - 100%),且在54份微生物学阴性标本中的50份未发现病原体(特异性93%,95%CI 82 - 98%)。单引物对PCR的敏感性为71%(44 - 90%),特异性为94%(85 - 99%)。传统染色方法(直接和间接荧光抗体及甲苯胺蓝 - O试验)的敏感性显著低于巢式PCR(38 - 53%)(p < 0.05)。在BAL液中,两种PCR方法均不比传统染色方法显著更好。在10例免疫功能低下患者的BAL液或痰中检测到卡氏肺孢子虫,但无卡氏肺孢子虫肺炎的微生物学证据,这表明可能存在无症状携带者或亚临床感染。在3例播散性肺孢子虫病患者中的2例血液中通过巢式PCR检测到卡氏肺孢子虫,但在仅局限于肺部的卡氏肺孢子虫感染的11例患者中仅1例检测到。对诱导痰进行巢式PCR在诊断卡氏肺孢子虫肺炎方面比传统染色方法更敏感,并提供了一种检测播散性疾病的非侵入性方法。