背景:由于艾滋病流行和免疫抑制疗法,真菌感染在过去几年中有所增加。可用的诊断方法,比如文化,组织病理学和血清学,在灵敏度方面有几个缺点,特异性和耗时,而分子方法仍然昂贵且依赖于许多设备。为了克服这些挑战,等温核酸扩增技术(INAT)成为有前途的传染病诊断方法。
目的:本文旨在介绍和讨论等温核酸扩增技术在医学真菌学中的主要贡献。
方法:包含每个INAT术语的论文(NASBA,RCA,LAMP,CPA,SDA,HAD或PSR)和术语“真菌病”或“疾病”,真菌是从国家生物技术信息中心数据库获得的,直到2019年8月。
结果:NASBA,RCA,LAMP和PSR是文献中报道的用于检测和鉴定病原真菌的INAT。尽管以前需要常规PCR,RCA技术也可能用于基因分型或隐蔽物种分化,这对某些真菌病的治疗可能很重要;尽管如此,LAMP是用于病原体检测的最常用的INAT。
结论:在本文审查的所有INAT中,LAMP似乎是最适合真菌检测的方法,因为它是负担得起的,敏感,具体,用户友好,快速,健壮,无设备且可交付给最终用户,符合世界卫生组织的所有公认标准,以获得理想的诊断方法。
BACKGROUND: Fungal infections have increased during the last years due to the AIDS epidemic and immunosuppressive therapies. The available diagnostic methods, such as culture, histopathology and serology, have several drawbacks regarding sensitivity, specificity and time-consuming, while molecular methods are still expensive and dependent on many devices. In order to overcome these challenges, isothermal nucleic acid amplification techniques (INAT) arose as promising diagnostic methods for infectious diseases.
OBJECTIVE: This
review aimed to present and discuss the main contributions of the isothermal nucleic acid amplification techniques applied in medical mycology.
METHODS: Papers containing terms for each INAT (NASBA, RCA, LAMP, CPA, SDA, HAD or PSR) and the terms \'mycoses\' or \'disease, fungal\' were obtained from National Center for Biotechnology Information database until August 2019.
RESULTS: NASBA, RCA, LAMP and PSR are the INAT reported in the literature for detection and identification of pathogenic fungi. Despite the need of a previous conventional PCR, the RCA technique might also be used for genotyping or cryptic species differentiation, which may be important for the treatment of certain mycoses; nevertheless, LAMP is the most used INAT for pathogen detection.
CONCLUSIONS: Among all INATs herein reviewed, LAMP seems to be the most appropriate method for fungal detection, since it is affordable, sensitive, specific, user-friendly, rapid, robust, equipment-free and deliverable to end-users, fulfilling all ASSURED criteria of the World Health Organization for an ideal diagnostic method.