Environmental Analysis Laboratory, Environmental Protection Administration, Zhongli, Taiwan.
BMC Infect Dis. 2011 Apr 13;11:91. doi: 10.1186/1471-2334-11-91.
Airborne fungi play an important role in causing allergy and infections in susceptible people. Identification of these fungi, based on morphological characteristics, is time-consuming, expertise-demanding, and could be inaccurate.
We developed an oligonucleotide array that could accurately identify 21 important airborne fungi (13 genera) that may cause adverse health problems. The method consisted of PCR amplification of the internal transcribed spacer (ITS) regions, hybridization of the PCR products to a panel of oligonucleotide probes immobilized on a nylon membrane, and detection of the hybridization signals with alkaline phosphatase-conjugated antibodies.
A collection of 72 target and 66 nontarget reference strains were analyzed by the array. Both the sensitivity and specificity of the array were 100%, and the detection limit was 10 pg of genomic DNA per assay. Furthermore, 70 fungal isolates recovered from air samples were identified by the array and the identification results were confirmed by sequencing of the ITS and D1/D2 domain of the large-subunit RNA gene. The sensitivity and specificity of the array for identification of the air isolates was 100% (26/26) and 97.7% (43/44), respectively.
Identification of airborne fungi by the array was cheap and accurate. The current array may contribute to decipher the relationship between airborne fungi and adverse health effect.
空气中的真菌在引起易感人群过敏和感染方面起着重要作用。基于形态特征对这些真菌进行鉴定既耗时，又需要专业知识，而且可能不准确。
我们开发了一种寡核苷酸阵列，可以准确识别可能导致健康问题的 21 种重要空气传播真菌（13 属）。该方法包括对内部转录间隔区（ITS）区域进行 PCR 扩增，将 PCR 产物与固定在尼龙膜上的一组寡核苷酸探针进行杂交，并用碱性磷酸酶缀合的抗体检测杂交信号。
通过该阵列分析了 72 个目标和 66 个非目标参考菌株。该阵列的灵敏度和特异性均为 100%，检测限为每个检测 10 pg 基因组 DNA。此外，还通过该阵列鉴定了从空气样本中回收的 70 株真菌分离株，鉴定结果通过 ITS 和大亚基 RNA 基因的 D1/D2 结构域的测序得到确认。该阵列对空气分离株的鉴定灵敏度和特异性分别为 100%（26/26）和 97.7%（43/44）。
通过该阵列鉴定空气中的真菌既便宜又准确。该阵列可能有助于阐明空气传播真菌与不良健康影响之间的关系。
