Microbiology Department, Kelvin Building, The Royal Group of Hospitals and Dental Hospital Health and Social Services Trust, Belfast, BT12 6BA, UK.
J Microbiol Methods. 2010 Mar;80(3):257-61. doi: 10.1016/j.mimet.2010.01.004. Epub 2010 Jan 14.
A novel microarray was constructed with DNA PCR product probes targeting species specific functional genes of nine clinically significant respiratory pathogens, including the Gram-positive organisms (Streptococcus pneumoniae, Streptococcus pyogenes), the Gram-negative organisms (Chlamydia pneumoniae, Coxiella burnetii Haemophilus spp., Legionella pneumophila, Moraxella catarrhalis, and Pseudomonas aeruginosa), as well as the atypical bacterium, Mycoplasma pneumoniae. In a "proof-of-concept" evaluation of the developed microarray, the microarray was compared with real-time PCR from 14 sputum specimens from COPD patients. All of the samples positive for bacterial species in real-time PCR were also positive for the same bacterial species using the microarray. This study shows that a microarray using PCR probes is a potentially useful method to monitor the populations of bacteria in respiratory specimens and can be tailored to specific clinical needs such as respiratory infections of particular patient populations, including patients with cystic fibrosis and bronchiectasis.
构建了一种新型微阵列,其 DNA PCR 产物探针针对九种临床重要呼吸道病原体的种特异性功能基因,包括革兰氏阳性菌(肺炎链球菌、酿脓链球菌)、革兰氏阴性菌(肺炎衣原体、贝纳柯克斯体、嗜血杆菌属、嗜肺军团菌、卡他莫拉菌和铜绿假单胞菌)以及非典型细菌肺炎支原体。在对开发的微阵列进行的“概念验证”评估中,将微阵列与来自 COPD 患者的 14 份痰标本的实时 PCR 进行了比较。使用实时 PCR 检测到的所有细菌阳性样本,在微阵列检测中也为同一细菌阳性。这项研究表明,使用 PCR 探针的微阵列是监测呼吸道标本中细菌种群的一种潜在有用方法,并且可以根据特定的临床需求进行定制,例如特定患者人群的呼吸道感染,包括囊性纤维化和支气管扩张症患者。