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从葡萄牙环境空气中采集的曲霉属物种-对依曲康唑耐药的烟曲霉狭义种的分子鉴定、抗真菌药敏性和 cyp51A 基因测序。

Aspergillus species collected from environmental air samples in Portugal-molecular identification, antifungal susceptibility and sequencing of cyp51A gene on A. fumigatus sensu stricto itraconazole resistant.

机构信息

Laboratory of Microbiology, Biological Sciences Department, Faculty of Pharmacy of University of Porto, Porto, Portugal.

Laboratory of Microbiology, Service of Clinical Pathology, Centro Hospitalar S. João EPE, Porto, Portugal.

出版信息

J Appl Microbiol. 2019 Apr;126(4):1140-1148. doi: 10.1111/jam.14217. Epub 2019 Feb 27.

Abstract

AIMS

Aspergillus sp. are ubiquitous saprophytic fungi and their conidia easily inhaled. This is particularly important in immunocompromised patients, more susceptible to developing invasive aspergillosis. In addition to A. fumigatus sensu stricto, cryptic species, many resistant to antifungal drugs, have been associated with invasive infections, making it important to assess their presence and diversity in different environments. Therefore, the aim of this study was to assess the presence, diversity and susceptibility to antifungal drugs of airborne fungi. Moreover, in azole-resistant A. fumigatus sensu stricto isolates the presence of underlying molecular mechanisms of resistance was investigated.

METHODS AND RESULTS

Eighty-four Aspergillus isolates were collected from the environment air in hospitals and the Water Treatment Plant. The use of molecular tools allowed to detect 12 different cryptic species, showing a prevalence of 21·4%. The majority of isolates (69·0%) belonged to A. fumigatus complex and 65·4% were A. fumigatus sensu stricto. Among these, 21·8% were resistant to itraconazole (ITZ), 38·2% to posaconazole and 87·3% to isavuconazole; none of them were resistant to voriconazole or amphotericin B. Sequencing of the cyp51A gene on the 12 A. fumigatus sensu stricto ITZ-resistant isolates revealed the presence of mutations.

CONCLUSION

Our study reports a large number of environmental-resistant Aspergillus species, including A. fumigatus sensu stricto that display an important role in invasive fungal infections. None of the environmental isolates showed mutations on cyp51A gene related to azole resistance.

SIGNIFICANT AND IMPACT OF THE STUDY

This study is the first assessment of molecular resistance mechanisms in A. fumigatus sensu stricto environmental isolates, in Portugal. Since TR34/L98H and TR46/Y121F/T289A cyp51A mutations were already reported in the clinical setting in Portugal (Monteiro et al. J Glob Antimicron Resist 13: 190-191, 2018; Pinto et al. Front Microbiol 9: 1656, 2018), and have been linked to environmental route, it is utmost importance to perform surveillance network for azole-resistant A. fumigatus.

摘要

目的

烟曲霉是一种无处不在的腐生真菌,其分生孢子很容易被吸入。这在免疫功能低下的患者中尤为重要,他们更容易发生侵袭性曲霉病。除了严格意义上的烟曲霉之外,许多对抗真菌药物具有耐药性的隐球菌种也与侵袭性感染有关,因此评估其在不同环境中的存在和多样性非常重要。因此,本研究旨在评估空气中真菌的存在、多样性和抗真菌药物敏感性。此外,还研究了唑类耐药的严格意义上的烟曲霉分离株中潜在的耐药分子机制。

方法和结果

从医院和水疗中心的环境空气中采集了 84 株烟曲霉分离株。使用分子工具可以检测到 12 种不同的隐球菌种,其流行率为 21.4%。大多数分离株(69.0%)属于烟曲霉复合体,其中 65.4%为严格意义上的烟曲霉。其中,21.8%对伊曲康唑(ITZ)耐药,38.2%对泊沙康唑耐药,87.3%对伊曲康唑耐药;它们均对伏立康唑和两性霉素 B 敏感。对 12 株严格意义上的烟曲霉 ITZ 耐药分离株的 cyp51A 基因进行测序显示存在突变。

结论

本研究报告了大量环境中存在的耐药曲霉种,包括严格意义上的烟曲霉,它们在侵袭性真菌感染中起着重要作用。环境分离株中均未发现与唑类耐药相关的 cyp51A 基因突变。

本研究的意义和影响

这是葡萄牙首次对严格意义上的烟曲霉环境分离株的分子耐药机制进行评估。由于葡萄牙已报道了 TR34/L98H 和 TR46/Y121F/T289A cyp51A 突变与临床相关(Monteiro 等人,J Glob Antimicron Resist 13:190-191,2018;Pinto 等人,Front Microbiol 9:1656,2018),且与环境途径有关,因此对唑类耐药的烟曲霉进行监测网络非常重要。

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