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数字 PCR——一种在微生物学领域具有广泛应用的新兴技术。

Digital PCR-An Emerging Technology with Broad Applications in Microbiology.

机构信息

Department of Laboratory Medicine, University of Washington, Seattle, WA.

Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA.

出版信息

Clin Chem. 2020 Jan 1;66(1):117-123. doi: 10.1373/clinchem.2019.304048.

Abstract

BACKGROUND

The PCR and its variant, quantitative PCR (qPCR), have revolutionized the practice of clinical microbiology. Continued advancements in PCR have led to a new derivative, digital PCR (dPCR), which promises to address certain limitations inherent to qPCR.

CONTENT

Here we highlight the important technical differences between qPCR and dPCR, and the potential advantages and disadvantages of each. We then review specific situations in which dPCR has been implemented in clinical microbiology and the results of such applications. Finally, we attempt to place dPCR in the context of other emerging technologies relevant to the clinical laboratory, including next-generation sequencing.

SUMMARY

dPCR offers certain clear advantages over traditional qPCR, but these are to some degree offset by limitations of the technology, at least as currently practiced. Laboratories considering implementation of dPCR should carefully weigh the potential advantages and disadvantages of this powerful technique for each specific application planned.

摘要

背景

PCR 及其变体,定量 PCR(qPCR),彻底改变了临床微生物学的实践。PCR 的持续进步导致了一种新的衍生技术,数字 PCR(dPCR),它有望解决 qPCR 固有的某些限制。

内容

在这里,我们强调了 qPCR 和 dPCR 之间的重要技术差异,以及每种方法的潜在优缺点。然后,我们回顾了 dPCR 在临床微生物学中的具体应用情况以及这些应用的结果。最后,我们试图将 dPCR 置于与临床实验室相关的其他新兴技术的背景下,包括下一代测序。

总结

dPCR 相对于传统的 qPCR 具有某些明显的优势,但这些优势在某种程度上被该技术的局限性所抵消,至少就目前的实践而言。考虑实施 dPCR 的实验室应根据每个特定应用计划仔细权衡该强大技术的潜在优缺点。

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