Use of subtractive hybridization to identify a diagnostic probe for a cystic fibrosis epidemic of Pseudomonas aeruginosa
Use of subtractive hybridization to identify a diagnostic probe for a cystic fibrosis epidemic of Pseudomonas aeruginosa
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DOI：
10.1128/jcm.40.12.4607-4611.2002
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发表时间：
2002-12-01
影响因子：
9.4
通讯作者：
Winstanley, C
中科院分区：
文献类型：
--
作者：
Parsons, YN;Panagea, S;Winstanley, C
一种多重耐药的铜绿假单胞菌菌株在英国利物浦诊所就诊的囊性纤维化(CF)患者中广泛存在。采用抑制性消减杂交技术，通过斑点杂交和聚合酶链式反应(PCR)扩增分析，鉴定了利物浦CF流行株中存在但不存在于PAO1株中的序列，确定了这些序列在一组CF分离株中的流行率。只在利物浦流行株中发现了几个序列。在利物浦流行株和少数其他分离株中存在一些序列，包括与O6血清型和铁载体产生有关的基因的同源性序列。利物浦流行株和81%的非流行分离株含有一个序列，被鉴定为PAGI-1基因组岛的一部分。其他与疫情传播有关的菌株来自英国曼彻斯特和澳大利亚墨尔本，也进行了筛查。在曼彻斯特菌株中没有发现任何已鉴定的序列。然而，两个墨尔本毒株中的一个含有利物浦流行毒株中的一些序列。所有涉及疫情传播的分离株和76%的零星分离株都含有exos基因。所有利物浦流行毒株的分离株都存在一个序列，用来开发一种诊断PCR试验，用于从菌落或直接从痰样本中鉴定该毒株。
A multiresistant strain of Pseudomonas aeruginosa is widespread among cystic fibrosis (CF) patients attending clinics in Liverpool, United Kingdom. Suppression subtractive hybridization was used to identify sequences present in the Liverpool CF epidemic strain but absent from strain PAO1 Using dot blot and PCR amplification assays, the prevalence of such sequences among a panel of CF isolates was determined. Several sequences were found only in the Liverpool epidemic strain. Some sequences were present in the Liverpool epidemic strain and in a minority of other isolates, including sequences with homology to genes implicated in O6 serotype and siderophore production. The Liverpool epidemic strain and 81% of nonepidemic isolates contained a sequence identified as part of the PAGI-1 genomic island. Other strains implicated in epidemic spread, which were from Manchester, United Kingdom, and Melbourne, Australia, were also screened. None of the sequences identified was present in the Manchester strain. However, one of two Melbourne strains contained some of the sequences found in the Liverpool epidemic strain. All isolates implicated in epidemic spread and 76% of sporadic isolates contained the exoS gene. A sequence present in all isolates of the Liverpool epidemic strain was used to develop a diagnostic PCR test for identification of the strain from colonies or directly from sputum samples.