基质辅助激光解吸电离飞行时间质谱分析技术在微生物鉴定与分型中的应用

微生物的鉴定方法有很多种, 这些方法基于表型、免疫学、遗传学等, 目前普遍采用16S rRNA和18S rRNA基因测序为标准, 但这些方法也有一些局限性, 费时费力、操作复杂、劳动力和试剂成本都很高, 限制了在常规微生物检测实验室的普及与推广。近年来, 基质辅助激光解吸电离飞行时间质谱法(matrix-assisted laser desorption ionization time-of-flight mass spectrometry, MALDI-TOF MS)在微生物学领域的应用越来越重要, 与应用于微生物鉴定的其他技术不同, MALDI-TOF MS更快速、准确、经济且易于操作。随着样品制备、数据库丰富和算法优化的发展, 使用MALDI-TOF MS进行鉴定的准确性和速度不断提高。虽然大多数研究都集中在细菌上, 但这项技术也应用于真菌和病毒。本文对MALDI-TOF MS技术对细菌、真菌、病毒鉴定与分型的研究进展以及目前该技术的局限性进行综述, 旨为该技术在临床诊断、食品检测等领域的多种应用奠定基础。

Application of matrix-assisted laser desorption ionization time-of-flight mass spectrometry in microbial identification and typing

There are many methods for microbial identification, which are based on phenotype, immunology, genetics, etc. At present, 16S rRNA and 18S rRNA gene sequencing are widely used as the standards, but these methods also have some limitations, such as time consuming, labor consuming, complex operation, high labor and reagent costs, which limit the popularization and promotion in routine microbiological testing laboratories. In recent years, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has become more and more important in the field of microbiology. Unlike other technologies used in microbial identification, MALDI-TOF MS is faster, more accurate, more economical and easier to operate. With the development of sample preparation, rich database and algorithm optimization, the accuracy and speed of identification by MALDI-TOF MS are increasing. Although most studies are focused on bacteria, this technology is also applied to fungi and viruses. This paper reviewed the advances in the identification and typing of bacteria, fungi and viruses using MALDI-TOF MS technology as well as the limitations of this technology, aiming to lay the foundation for its multiple applications in clinical diagnosis, food detection and other fields.