Gillespie S H, Ullman C, Smith M D, Emery V
Division of Communicable Diseases, Royal Free Hospital School of Medicine, London, United Kingdom.
J Clin Microbiol. 1994 May;32(5):1308-11. doi: 10.1128/jcm.32.5.1308-1311.1994.
A method for the detection of Streptococcus pneumoniae in sputum samples by PCR has been developed. The assay employs oligonucleotide primers specific for a portion of the autolysin gene lytA of S. pneumoniae. Other closely related streptococci, Haemophilus influenzae, and Moraxella catarrhalis do not give a positive result in the assay. The assay was capable of detecting between 10 and 100 CFU of S. pneumoniae in distilled water and 1.4 x 10(4) CFU/ml in simulated sputum samples. Sputum samples from 33 patients with acute pneumonia were collected and subjected to culture, PCR, and C-polysaccharide antigen detection by enzyme-linked immunosorbent assay (ELISA). A significant isolate of S. pneumoniae was isolated from 14 patients, of which 13 were positive by PCR and C-polysaccharide antigen ELISA. No positive results were obtained for the 19 patients in whom other pathogens or upper respiratory tract floras only were isolated. The sensitivity of the autolysin PCR is 92.8%, the specificity is 100%, the predictive value of a positive result is 100%, and the predictive value of a negative result is 95%. This suggests that autolysin PCR is suitable for the detection of S. pneumoniae in clinical samples.
已开发出一种通过聚合酶链反应(PCR)检测痰标本中肺炎链球菌的方法。该检测方法采用了针对肺炎链球菌自溶素基因lytA一部分的寡核苷酸引物。其他密切相关的链球菌、流感嗜血杆菌和卡他莫拉菌在该检测中不会产生阳性结果。该检测方法能够在蒸馏水中检测到10至100个肺炎链球菌菌落形成单位(CFU),在模拟痰标本中检测到1.4×10⁴CFU/ml。收集了33例急性肺炎患者的痰标本,并进行培养、PCR以及通过酶联免疫吸附测定(ELISA)检测C多糖抗原。从14例患者中分离出显著的肺炎链球菌菌株,其中13例通过PCR和C多糖抗原ELISA检测呈阳性。在仅分离出其他病原体或上呼吸道菌群的19例患者中未获得阳性结果。自溶素PCR的敏感性为92.8%,特异性为100%,阳性结果预测值为100%,阴性结果预测值为95%。这表明自溶素PCR适用于临床标本中肺炎链球菌的检测。