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通过聚合酶链反应-限制性片段长度多态性(PCR-RFLP)和聚合酶链反应-真菌特异性引物(PCR-FSP)方法对从指甲标本中分离出的酵母群落进行分子鉴定。

Molecular identification of yeast communities isolated from nail specimens by PCR-RFLP and PCR-FSP methods.

作者信息

Jabrodini Ahmad, Zaighami Mitra, Khodadadi Ali, Pakshir Keyvan, Nouraei Hasti, Khodadadi Hossein

机构信息

Department of Medical Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.

Faculty of Medicine, Najafabad Branch, Islamic Azad University, Najafabad, Isfahan, Iran.

出版信息

Curr Med Mycol. 2024 May 7;10:e2024.345184.1539. doi: 10.22034/cmm.2024.345237.1539. eCollection 2024.

Abstract

BACKGROUND AND PURPOSE

Onychomycosis is a common fungal infection that affects the nails, caused by various fungal agents. Moreover, yeast onychomycosis has increased in recent years. Yeast isolates might not be identified at the species level by conventional methods, whereas molecular methods can identify yeast isolates more accurately. This study aimed to identify yeast communities isolated from nail specimens by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) and PCR- fragment size polymorphism (FSP) methods.

MATERIALS AND METHODS

This experimental study was conducted on archival yeast isolates obtained from 269 patients suspected of onychomycosis who referred to the Medical Mycology Laboratory at Shiraz University of Medical Sciences in Shiraz, Iran, between April 2022 and March 2023. Onychomycosis was confirmed through direct examination and culture of nail specimens. The PCR-RFLP and PCR-FSP methods were used to identify yeast isolates.

RESULTS

In total, 78 (28.99%) yeast strains were identified. was the most common species, followed by complex and . Uncommon species of yeasts, such as , , , and were identified by molecular methods. The PCR-FSP method showed a strong agreement with the PCR-RFLP method in the identification of common yeast agents causing onychomycosis (κ=0.84).

CONCLUSION

It seems necessary to use molecular diagnostic tools in addition to conventional methods to identify yeast isolates in clinical laboratories. The rapid and accurate identification of fungal agents causing onychomycosis is useful for the selection of an appropriate treatment strategy.

摘要

背景与目的

甲癣是一种常见的真菌感染,由多种真菌病原体引起,影响指甲。此外,近年来酵母菌性甲癣有所增加。传统方法可能无法在种水平上鉴定酵母菌分离株,而分子方法可以更准确地鉴定酵母菌分离株。本研究旨在通过聚合酶链反应(PCR)-限制性片段长度多态性(RFLP)和PCR-片段大小多态性(FSP)方法鉴定从指甲标本中分离出的酵母菌群落。

材料与方法

本实验研究对2022年4月至2023年3月期间转诊至伊朗设拉子医科大学医学真菌实验室的269例疑似甲癣患者的存档酵母菌分离株进行。通过指甲标本的直接检查和培养确诊甲癣。采用PCR-RFLP和PCR-FSP方法鉴定酵母菌分离株。

结果

共鉴定出78株(28.99%)酵母菌菌株。 是最常见的菌种,其次是 复合体和 。通过分子方法鉴定出了罕见的酵母菌菌种,如 、 、 和 。在鉴定引起甲癣的常见酵母菌病原体方面,PCR-FSP方法与PCR-RFLP方法显示出高度一致性(κ=0.84)。

结论

临床实验室似乎有必要在传统方法之外使用分子诊断工具来鉴定酵母菌分离株。快速准确地鉴定引起甲癣的真菌病原体有助于选择合适的治疗策略。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8adb/11688582/998c8561fab9/CMM-10-e2024.345184.1539-g001.jpg

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