Department of Medical Sciences, Section of Clinical Microbiology, Uppsala University, Uppsala, Sweden; Department of Medical Sciences, Section of Infectious Diseases, Uppsala University, Uppsala, Sweden; Centre of Clinical Research, Falu Hospital, Falun, Sweden.
Department of Medical Sciences, Section of Clinical Microbiology, Uppsala University, Uppsala, Sweden.
J Mycol Med. 2019 Apr;29(1):1-6. doi: 10.1016/j.mycmed.2019.01.009. Epub 2019 Feb 11.
Dermatophytes are common pathogens in superficial mycoses that are routinely identified by culture or PCR analysis of freshly collected skin, nail or hair specimens. Although clinical samples are normally processed without delay, practical or research issues may necessitate sample storage until later analysis. However, the influence of extended sample storage on the ability to recover fungi by culture vs. PCR analysis has not been specifically studied. Here, a total of 172 dermatological samples collected from 2013-2015 were examined before and after refrigerated storage at 4°C for 10.2-32.3 (mean 25.6) months. By culture, 35% of the dermatophyte-containing fresh samples remained positive at re-examination. At species level, only 19% of initially Trichophyton rubrum-positive samples yielded a positive result after refrigeration, whereas few samples containing Trichophyton violaceum, Microsporum canis or Microsporum audouinii remained culture-positive. Using PCR, 76% of dermatophyte DNA-positive fresh samples were still positive at re-analysis. Notably, 92% of the samples targeted by the T. rubrum DNA primer remained positive after storage. Hence, PCR analysis is more favourable than cultivation with regard to the detectability of dermatophytes in long-term refrigerated clinical samples.
皮肤癣菌是常见的浅部真菌病病原体,通常通过对新鲜采集的皮肤、指甲或头发标本进行培养或 PCR 分析来鉴定。虽然临床标本通常会立即进行处理,但实际或研究问题可能需要将标本储存起来,以便以后进行分析。然而,长时间储存标本对培养和 PCR 分析检测真菌能力的影响尚未得到专门研究。在这里,总共对 172 份于 2013-2015 年采集的皮肤科标本进行了研究,这些标本在 4°C 下冷藏储存了 10.2-32.3(平均 25.6)个月后,进行了冷藏前后的检查。通过培养,35%的含皮肤癣菌的新鲜标本在重新检查时仍然呈阳性。在种水平上,最初 Trichophyton rubrum 阳性的标本中只有 19%在冷藏后产生了阳性结果,而含有 Trichophyton violaceum、Microsporum canis 或 Microsporum audouinii 的少数标本仍保持培养阳性。通过 PCR,76%的含皮肤癣菌 DNA 阳性的新鲜标本在重新分析时仍然呈阳性。值得注意的是,92%的 T. rubrum DNA 引物靶向的标本在储存后仍保持阳性。因此,与培养相比,PCR 分析在长期冷藏的临床标本中更有利于检测皮肤癣菌。