Specific and rapid detection by fluorescent in situ hybridization of bacteria in clinical samples obtained from cystic fibrosis patients
Specific and rapid detection by fluorescent in situ hybridization of bacteria in clinical samples obtained from cystic fibrosis patients
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DOI：
10.1128/jcm.38.2.818-825.2000
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发表时间：
2000-02-01
影响因子：
9.4
通讯作者：
Heesemann, J
中科院分区：
文献类型：
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作者：
Hogardt, M;Trebesius, K;Heesemann, J
我们报告了通过荧光原位杂交 (FISH) 对囊性纤维化 (CF) 患者临床标本中常见的细菌进行快速、特异检测，在比较序列分析的基础上，我们设计了与这些微生物的物种特异性 16S rRNA 区域互补的寡核苷酸探针，并通过不同远缘相关以及不同细菌的杂交证明了探针的特异性。 密切相关的参考菌株。此外，在一个试点项目中，我们通过 FISH 研究了 CF 患者的 75 份痰样本和 10 份咽拭子样本，并在这些样本中检测到了铜绿假单胞菌、洋葱伯克霍尔德菌、嗜麦芽寡养单胞菌、流感嗜血杆菌和金黄色葡萄球菌。与传统微生物培养的结果相比，FISH 的特异性为 100%。相比之下，标准实验室培养的灵敏度稍高，因为 FISH 对痰样本中细菌的显微镜检测极限约为 4 x 10(5) CFU/ml 痰（导致 FISH 的灵敏度为 90%）。此外，我们证明 FISH 将有助于快速检测导致 CF 患者急性肺部病情加重的细菌，正如在流感嗜血杆菌、金黄色葡萄球菌和铜绿假单胞菌病情加重患者中所证明的那样。因此，FISH 是快速、特异性检测 CF 患者（尤其是肺部病情加重患者）临床样本中细菌的一种有价值的附加方法。
We report on the rapid and specific detection of bacteria commonly isolated from clinical specimens from cystic fibrosis (CF) patients by fluorescent in situ hybridization (FISH), On the basis of comparative sequence analysis, we designed oligonucleotide probes complementary to species-specific 16S rRNA regions of these microorganisms and demonstrated the specificities of the probes by hybridization of different remotely related as well as closely related reference strains. Furthermore, in a pilot project we investigated 75 sputum samples and 10 throat swab specimens from CF patients by FISH and detected Pseudomonas aeruginosa, Burkholderia cepacia, Stenotrophomonas maltophilia, Haemophilus influenzae, and Staphylococcus aureus within these specimens. The specificity of FISH was 100% in comparison to the results of conventional microbial culture. In contrast, the sensitivity of standard laboratory cultivation was moderately higher, since the limit for microscopic detection of bacteria within sputum samples by FISH was approximately 4 x 10(5) CFU/ml of sputum (resulting in a 90% sensitivity for FISH). Moreover, we demonstrated that FISH will be useful for the rapid detection of bacteria that cause acute pulmonary exacerbations in CF patients, as demonstrated in patients with H. influenzae, S. aureus, and P. aeruginosa exacerbations. Therefore, FISH is a valuable additional method for the rapid and specific detection of bacteria in clinical samples from CF patients, in particular, patients with pulmonary exacerbations.