嗜肺军团菌15种血清型基因芯片检测方法的建立

摘要

To establish a rapid, accurate detection method for 15 distinct O antigen forms of Legionella pneumophila using DNA microarray combined with multiplex PCR. The special genes of these 15 distinct O antigen forms of L. Pneumophila were as target genes for multiplex PCR respectively, then primers and captured oligonucleotide probes were hybridizes with DNA microarray, which contained specific probes of Legionella pneumophila. Scanner was used to determinant the types of bacterium. The DNA microarray assay can detect 15 distinct O antigen forms of L. Pneumophila specially. The sensitivity of the DNA microarray was 10 ng. The detection method of 15 distinct O antigen forms of Legionella pneumophila by DNA microarray was established. The detection method has better specificity, sensitivity and repeatability.%建立一种多重PCR技术结合基因芯片检测方法，实现嗜肺军团菌15种血清型的快速、准确检测。根据嗜肺军团菌15种血清型的O抗原特异性基因设计并筛选合适的引物和探针，进行多重PCR扩增，制备寡核苷酸芯片。将多重PCR扩增产物与带有特异探针的芯片杂交。用扫描仪扫描，判定嗜肺军团菌的血清型。该基因芯片可特异性的检测嗜肺军团菌的15种血清型，具有良好的特异性，芯片纯菌DNA检测灵敏度为10 ng。所建立的嗜肺军团菌15种血清型基因芯片检测方法特异性好，灵敏度高，具有较好的实用性。