Dipartimento di Sanità Pubblica e Zootecnia, Università degli Studi di Bari, Bari, Italy.
Med Mycol. 2013 Feb;51(2):136-43. doi: 10.3109/13693786.2012.691995. Epub 2012 Jun 11.
The few studies attempting to specifically characterize dermatophytes from hair samples of dogs and cats using PCR-based methodology relied on sequence-based analysis of selected genetic markers. The aim of the present investigation was to establish and evaluate a PCR-based approach employing genetic markers of nuclear DNA for the specific detection of dermatophytes on such specimens. Using 183 hair samples, we directly compared the test results of our one-step and nested-PCR assays with those based on conventional microscopy and in vitro culture techniques (using the latter as the reference method). The one step-PCR was highly accurate (AUC > 90) for the testing of samples from dogs, but only moderately accurate (AUC = 78.6) for cats. A nested-PCR was accurate (AUC = 93.6) for samples from cats, and achieved higher specificity (94.1 and 94.4%) and sensitivity (100 and 94.9%) for samples from dogs and cats, respectively. In addition, the nested-PCR allowed the differentiation of Microsporum canis from Trichophyton interdigitale (zoophilic) and geophilic dermatophytes (i.e., Microsporum gypseum or Trichophyton terrestre), which was not possible using the one step-assay. The PCRs evaluated here provide practical tools for diagnostic applications to support clinicians in initiating prompt and targeted chemotherapy of dermatophytoses.
本研究旨在建立并评估一种基于聚合酶链反应(PCR)的方法,该方法采用核 DNA 的遗传标记,用于对这些标本进行特定的皮肤癣菌检测。使用 183 份毛发样本,我们将一步法和巢式 PCR 检测的结果与传统显微镜检查和体外培养技术(以后者作为参考方法)的检测结果进行了直接比较。对于犬的样本,一步法 PCR 的检测结果非常准确(AUC>90),但对于猫的样本仅为中度准确(AUC=78.6)。对于猫的样本,巢式 PCR 检测结果准确(AUC=93.6),对于犬和猫的样本,分别具有更高的特异性(94.1%和 94.4%)和敏感性(100%和 94.9%)。此外,巢式 PCR 可区分犬小孢子菌(亲动物性)和亲土性皮肤癣菌(即石膏样小孢子菌或须毛癣菌),而一步法无法实现这种区分。这里评估的 PCR 为诊断应用提供了实用工具,以支持临床医生及时、有针对性地开展皮肤癣菌病的化疗。