Nieguitsila Adélaïde, Deville Manjula, Jamal Taoufik, Halos Lénaïg, Berthelemy Madeleine, Chermette René, Latouche Sophie, Arné Pascal, Guillot Jacques
INRA, AFSSA, ENVA, UPVM, UMR 956, Ecole Nationale Vétérinaire d'Alfort, Maisons-Alfort, France.
J Microbiol Methods. 2007 Jul;70(1):86-95. doi: 10.1016/j.mimet.2007.03.021. Epub 2007 Apr 12.
Information obtained from fungal air samples can assist in the assessment of health hazards and can be useful in proactive indoor air quality monitoring. The objective of the present study was to evaluate the PCR-TTGE technique for the analysis of fungal diversity in the air. Eleven air samples were collected in five different sites using the bioimpactor CIP 10-M (Arelco). After a 2 hours sampling period, the collection liquid was recovered for subsequent cultivation and PCR-TTGE. A set of three fungi-specific primers (Fungcont 1, Fungcont 2+GC and Fungcont 3) was designed for the partial amplification of the 18S rRNA gene. The amplification was obtained in a single reaction tube by a semi-nested PCR. For identification, the TTGE bands were extracted and sequenced. PCR-TTGE allowed the clear separation of amplicons corresponding to distinct fungal species (both Ascomycota and Basidiomycota) that may be encountered in air. The number of fungal taxa detected after culture was systematically higher than the number of taxa found using PCR-TTGE. However, few fungal species were detected by PCR-TTGE and not by cultivation, suggesting that the combination of these approaches may provide a better analysis of fungal diversity in air samples than either method alone.
从真菌空气样本中获取的信息有助于评估健康危害，并且在室内空气质量的主动监测中很有用。本研究的目的是评估用于分析空气中真菌多样性的PCR-TTGE技术。使用生物冲击器CIP 10-M（Arelco）在五个不同地点采集了11个空气样本。在2小时的采样期后，回收收集液用于后续培养和PCR-TTGE。设计了一组三种真菌特异性引物（Fungcont 1、Fungcont 2+GC和Fungcont 3）用于18S rRNA基因的部分扩增。通过半巢式PCR在单个反应管中获得扩增产物。为了进行鉴定，提取并测序TTGE条带。PCR-TTGE能够清晰地分离与空气中可能遇到的不同真菌物种（子囊菌门和担子菌门）相对应的扩增子。培养后检测到的真菌分类单元数量系统地高于使用PCR-TTGE发现的分类单元数量。然而，通过PCR-TTGE检测到但未通过培养检测到的真菌物种很少，这表明这些方法的组合可能比单独使用任何一种方法都能更好地分析空气样本中的真菌多样性。
