Noriki Sakon, Ishida Hisaya
Department of Pathology, University of Fukui, Fukui, Japan
Ishida Dermatology Clinic, Fukui, Japan.
Med Mycol. 2016 Nov 1;54(8):808-15. doi: 10.1093/mmy/myw037. Epub 2016 Jun 1.
Tinea refers to superficial infection with one of three fungal genera-Microsporum, Epidermophyton, or Trichophyton-that are collectively known as dermatophytes. These infections are among the most common diseases worldwide and cause chronic morbidity. They are usually diagnosed by direct microscopy and fungal culture, which are burdensome to perform in the clinical setting. To supplement conventional methods, we developed a new method that employs an immunochromatography test for detection of dermatophyte infections. First, anti-Trichophyton monoclonal antibodies (mAb) were produced in mice using a Trichophyton allergen solution as an immunogen. The mAb specificity was assessed by immunostaining alcohol fixed slide cultures and formalin fixed paraffin-embedded microbial samples. Both alcohol- and formalin-fixed samples of all seven species of Trichophyton tested displayed positive immunostaining. Immunochromatography test strips were created using the anti-Trichophyton mAb. The efficiency of the test strip was assessed in patients diagnosed with tinea unguium and in healthy volunteers. Of the 20 patient nails tested, 19 tested positive and one tested negative, whereas of the 17 volunteer nails, only one tested positive. However, KOH microscopic examination of the volunteer nail that tested positive revealed the existence of Trichophyton hyphae. Although the number of nails assayed was small, since the assay had a sensitivity of 95.0% (19/20) and a specificity of 94.1% (16/17), the obtained results were considered to be promising. Thus, while further investigation with a greater number of samples is necessary, this method could potentially be employed as a new diagnostic tool for Trichophyton in the future.
癣是指由三种真菌属——小孢子菌属、表皮癣菌属或毛癣菌属——中的一种引起的浅表感染,这三种真菌统称为皮肤癣菌。这些感染是全球最常见的疾病之一,并会导致慢性发病。它们通常通过直接显微镜检查和真菌培养来诊断,在临床环境中进行这些检查很繁琐。为了补充传统方法,我们开发了一种新方法,该方法采用免疫层析试验来检测皮肤癣菌感染。首先,以毛癣菌变应原溶液作为免疫原,在小鼠体内产生抗毛癣菌单克隆抗体(mAb)。通过对酒精固定的载玻片培养物和福尔马林固定的石蜡包埋微生物样本进行免疫染色来评估单克隆抗体的特异性。所有测试的七种毛癣菌的酒精固定和福尔马林固定样本均显示阳性免疫染色。使用抗毛癣菌单克隆抗体制备免疫层析试纸条。在被诊断为甲癣的患者和健康志愿者中评估试纸条的效率。在测试的20个患者指甲中,19个呈阳性,1个呈阴性,而在17个志愿者指甲中,只有1个呈阳性。然而,对呈阳性的志愿者指甲进行氢氧化钾显微镜检查发现存在毛癣菌菌丝。尽管检测的指甲数量较少,但由于该检测的灵敏度为95.0%(19/20),特异性为94.1%(16/17),因此获得的结果被认为很有前景。因此,虽然需要用更多的样本进行进一步研究,但这种方法将来有可能作为一种新的毛癣菌诊断工具。