Diagnostic Performance of Direct Microscopic Techniques and Culture Media for Fungal Isolates from Hair Specimens

Faiza Zeeshan, F. Uddin, Suresh Kumar, Shahida Kashif, N. Jaffar
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Abstract

Objective: To compare the diagnostic performance of direct microscopic techniques and culture media for detecting fungal isolates in hair specimens.Study Design:  Cross-sectional study.Place and Duration of Study: The study was conducted at the Department of Microbiology, Basic Medical Sciences Institute, Karachi, Pakistan, from January 2021 to June 2021.Materials and Methods:  A total of 207 hair specimens were collected from the patients with superficial mycoses of the scalp. The specimens were collected under aseptic techniques using clean glass slides and forceps which were processed for Potassium hydroxide mount and Calcofluor white staining. The mycological cultures were performed on a Dermatophyte test medium and Sabouraud Dextrose agar with and without antibiotics. The data was recorded and analyzed on SPSS version 21. The frequencies, percentages, means, and ratios were calculated using Descriptive statistics. The association between variables were analyzed by the Chi-square test and kappa index.Results: Potassium hydroxide mount revealed fungal spores and hyphae in 154(74.3%) specimens, while Calcofluor white staining showed fungal elements in 179(86.5%) specimens. Out of 207 specimens, 106(51.2%) were culture positive and 101(48.8%) were declared culture negative. Out of culture-positive specimens, dermatophytes were yielded in 61(57.5%) and non-dermatophytes in 45(42.5%) specimens. Among all culture media, the highest number of dermatophytes was yielded by plain Sabouraud dextrose agar (59; 96.5%), followed by dermatophyte test medium (55; 90%), and Sabouraud dextrose agar with antibiotics (54; 90.1%). On the contrary, the primary isolation of non-dermatophytes on the plain was Sabouraud dextrose agar s 32(71.1%), Sabouraud dextrose agar with antibiotics 31(68.2%), and dermatophyte test medium 13(28.8%).Conclusion: In our study, we found Calcofluor white staining technique as the most sensitive method to detect fungal elements from the given hair specimens. For mycological culture, all tested culture media were equally effective for the retrieval of dermatophytes. However, for non-dermatophytes, plain SDA and SDA with antibiotics were found to be better media for isolation.
直接显微技术和培养基对头发标本真菌分离株的诊断性能
目的:比较直接显微技术和培养基对头发标本中分离真菌的诊断效果。研究设计:横断面研究。研究地点和时间:研究于2021年1月至2021年6月在巴基斯坦卡拉奇基础医学研究所微生物学系进行。材料与方法:采集头皮浅表性真菌病患者毛发标本207份。标本在无菌技术下采集,使用干净的玻璃载玻片和镊子,处理后进行氢氧化钾固定和氟化钙白色染色。真菌学培养分别在皮肤癣菌试验培养基和沙伯劳德葡萄糖琼脂上进行,有和没有抗生素。使用SPSS 21对数据进行记录和分析。使用描述性统计计算频率、百分比、平均值和比率。采用卡方检验和kappa指数分析变量间的相关性。结果:氢氧化钾涂片检出真菌孢子和菌丝154份(74.3%),荧光白染色检出真菌成分179份(86.5%)。207份标本中培养阳性106份(51.2%),培养阴性101份(48.8%)。在培养阳性标本中,61例(57.5%)产生皮肤真菌,45例(42.5%)产生非皮肤真菌。在所有培养基中,普通沙伯罗德葡萄糖琼脂产生的皮癣菌数量最多(59;96.5%),其次是皮菌试验培养基(55;90%),沙伯劳德葡萄糖琼脂加抗生素(54;90.1%)。与此相反,平原上非皮癣菌的主要分离株为Sabouraud葡萄糖琼脂32株(71.1%)、含抗生素的Sabouraud葡萄糖琼脂31株(68.2%)和皮癣菌试验培养基13株(28.8%)。结论:在我们的研究中,我们发现钙荧光白染色技术是最敏感的方法,从给定的头发标本中检测真菌元素。对于真菌学培养,所有测试的培养基对皮肤真菌的回收都是同样有效的。而对于非皮肤真菌,普通SDA和含抗生素的SDA是较好的分离培养基。
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