R&D Laboratory of Quality Control Material, Shanghai Center for Clinical Laboratory, Shanghai, China.
Shanghai Academy of Experimental Medicine, Shanghai, China.
Front Cell Infect Microbiol. 2023 Mar 17;13:1128337. doi: 10.3389/fcimb.2023.1128337. eCollection 2023.
Proficiency testing based on quality control materials is an important component of the quality assurance system for detection methods. However, in the detection of infectious diseases, it is a challenge to use quality control materials derived from clinical samples or pathogens owing to their infectious nature. The Xpert MTB/RIF assay, endorsed by the World Health Organization, is one of the most widely implemented assays in the detection of along with rifampicin resistance and its heterogeneity. Clinical isolates are typically used as quality controls for this assay, leading to concerns about biosafety, constrained target sequence polymorphisms, and time-consuming preparation. In this study, a heterogeneous quality control library for the Xpert MTB/RIF assay was constructed based on DNA synthesis and site-directed mutation, which provides sufficient rifampicin resistance polymorphisms, enabling monitoring all five probes of Xpert MTB/RIF and its combinations. and were used as heterogeneous hosts rather than the pathogen itself to eliminate biosafety risks; thus, preparation does not require a biosafety level III laboratory and the production time is reduced from a few months to a few days. The panel was stable for more than 15 months stored at 4°C and could be distributed at room temperature. All 11 laboratories in Shanghai participating in a pilot survey identified the specimens with corresponding probe patterns, and discordant results highlighted inappropriate operations in the process. Collectively, we show, for the first time, that this library, based on heterogeneous hosts, is an appropriate alternative for detection.
基于质控品的能力验证是检测方法质量保证体系的重要组成部分。然而，在传染病检测中，由于其感染性质，使用源自临床样本或病原体的质控品是一项挑战。世界卫生组织推荐的 Xpert MTB/RIF 检测是最广泛应用的检测方法之一，用于检测结核分枝杆菌和利福平耐药性及其异质性。该检测通常使用临床分离株作为质控品，这引发了对生物安全、有限的靶序列多态性和耗时的制备过程的关注。在这项研究中，我们基于 DNA 合成和定点突变构建了 Xpert MTB/RIF 检测的异质质控文库，该文库提供了足够的利福平耐药性多态性，能够监测 Xpert MTB/RIF 的所有五个探针及其组合。我们使用异质宿主而不是病原体本身作为异质宿主，以消除生物安全风险；因此，制备不需要三级生物安全实验室，并且生产时间从几个月缩短到几天。该面板在 4°C 下储存超过 15 个月仍然稳定，并且可以在室温下分发。上海参与试点调查的 11 家实验室均能识别出具有相应探针模式的标本，不一致的结果突出了过程中操作不当的问题。总之，我们首次表明，这种基于异质宿主的文库是一种合适的替代物，可用于结核分枝杆菌的检测。
