一次实时定量 PCR 检测铜绿假单胞菌，以区分囊性纤维化患者的间歇性感染和慢性感染。
One time quantitative PCR detection of Pseudomonas aeruginosa to discriminate intermittent from chronic infection in cystic fibrosis.
机构信息
Department of Infectious Disease, Medical Microbiology and Hygiene, University Hospital Heidelberg, Germany; Translational Lung Research Center Heidelberg (TLRC), German Center for Lung Research (DZL), University of Heidelberg, Heidelberg, Germany.
Department of Infectious Disease, Medical Microbiology and Hygiene, University Hospital Heidelberg, Germany.
出版信息
J Cyst Fibros. 2018 May;17(3):348-355. doi: 10.1016/j.jcf.2017.12.007. Epub 2018 Jan 12.
BACKGROUND
Chronic airway infection with Pseudomonas aeruginosa is a major risk factor of progression of lung disease in patients with cystic fibrosis (CF). Chronic P. aeruginosa infection evolves from intermittent infection that is amenable to antibiotic eradication, whereas chronically adapted P. aeruginosa becomes resistant to antibiotic therapy. Discrimination of intermittent versus chronic infection is therefore of high therapeutic relevance, yet the available diagnostic methods are only partly satisfactory. The aim of the present study was, therefore, to evaluate the usage of quantitative PCR (qPCR) to measure pathogen abundance and to discriminate between intermittent and chronic Pseudomonas infection in patients with CF.
METHOD
Using an established qPCR protocol, we analyzed the abundance of P. aeruginosa in 141 throats swabs and 238 sputa from CF patients with intermittent or chronic infection with P. aeruginosa, as determined by standard culture based diagnostics.
RESULTS
We observed a large increase of abundance of P. aeruginosa in throat swabs and sputum samples from patients with chronic compared to intermittent infections with P. aeruginosa. The data show that abundance of P. aeruginosa as measured by qPCR is a valuable tool to discriminate intermittent from chronic infection. Of note, P. aeruginosa burden seems more sensitive than mucoidity phenotype to discriminate chronic from intermittent strains. Furthermore we observed that molecular detection in throat swabs was linked to a viable culture in the sputum when sputum was available. This result is of special interest in young patients with cystic fibrosis that often cannot expectorate sputum. We also observed that qPCR in comparison to culture detected the infection earlier.
CONCLUSION
The results suggest that qPCR detection and quantification of P. aeruginosa is a precious tool to be added to the diagnostic toolbox in cystic fibrosis.
背景
铜绿假单胞菌（Pseudomonas aeruginosa）的慢性气道感染是囊性纤维化（cystic fibrosis，CF）患者肺部疾病进展的主要危险因素。慢性铜绿假单胞菌感染源自间歇性感染，这种感染可通过抗生素清除，而慢性适应的铜绿假单胞菌则对抗生素治疗产生耐药性。因此，区分间歇性感染和慢性感染具有重要的治疗意义，但现有的诊断方法并不完全令人满意。本研究旨在评估定量聚合酶链反应（qPCR）在 CF 患者中用于测量病原体丰度和区分间歇性与慢性铜绿假单胞菌感染的用途。
方法
使用既定的 qPCR 方案，我们分析了 141 例 CF 患者的咽喉拭子和 238 例慢性或间歇性铜绿假单胞菌感染患者的痰液中 P. aeruginosa 的丰度，这些患者的感染情况是通过基于标准培养的诊断方法确定的。
结果
我们观察到慢性感染患者的咽喉拭子和痰液样本中 P. aeruginosa 的丰度明显高于间歇性感染患者。数据表明，qPCR 测量的 P. aeruginosa 丰度是区分间歇性与慢性感染的有用工具。值得注意的是，与黏液表型相比，P. aeruginosa 负担更能敏感地区分慢性与间歇性菌株。此外，我们观察到当有痰样时，咽喉拭子的分子检测与痰中的培养物有关。这一结果在经常无法咳出痰液的年轻 CF 患者中特别有意义。我们还观察到 qPCR 与培养相比更早地检测到感染。
结论
这些结果表明，qPCR 检测和定量 P. aeruginosa 是 CF 诊断工具包中增加的宝贵工具。
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