Background

Parasitic infections are responsible for a significant burden of disease worldwide as a result of international travel and immigration. More accurate diagnostic tools are necessary in support to parasite control and elimination programmes in endemic regions as well as for rapid case detection in non-endemic areas. Digital PCR (dPCR) is a powerful technology with recent applications in parasitology.

Aims

This review provides for the first time an overview of dPCR as a novel technology applied to detection of parasitic infections, and highlights the most relevant potential benefits of this assay.

Sources

Peer-reviewed literature pertinent to this review based on PubMed, Cochrane and Embase databases as well as laboratory experience of authors.

Content

Among the 86 studies retrieved, 17 used the dPCR applied to parasites belonging to protozoa (8), helminths (8) and arthropods (1) of clinical human interest. dPCR was adopted in four studies, respectively, for Plasmodium and Schistosoma japonicum. dPCR led to clear advantages over quantitative real-time PCR in P. falciparum and spp., and in S. japonicum showing higher sensitivity; and in Cryptosporidium with higher stability to inhibitors from stool. For all parasites, dPCR allows absolute quantitation without the need of a standard curve. Various dPCR platforms were used. A few critical factors need consideration: DNA load, choice of platform and reaction optimization.

Implications

Owing to its sensitivity and quantitative characteristics, dPCR is a potential candidate to become an appealing new method among the molecular technologies for parasite detection and quantitative analysis in the future. In general, it has more applications than genomic DNA detection only, such as quantitation in mixed infections, gene expression and mutation analysis. dPCR should be considered in malaria screening and diagnosis as a complement to routine assays and in schistosomiasis elimination programmes. Standardized strategies and further studies are needed for the integration of dPCR in routine clinical laboratory.

中文翻译：

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数字PCR：诊断寄生虫感染的新技术

背景

由于国际旅行和移民，寄生虫感染在世界范围内造成了巨大的疾病负担。为了支持流行地区的寄生虫控制和消除计划以及非流行地区的快速病例检测，必须使用更准确的诊断工具。数字PCR（dPCR）是一项功能强大的技术，最近在寄生虫学中得到了应用。

目的

这篇综述首次提供了dPCR的概述，它是一种用于检测寄生虫感染的新技术，并着重介绍了此测定法最相关的潜在益处。

资料来源

基于PubMed，Cochrane和Embase数据库以及作者的实验室经验，与该评论相关的同行评审文献。

内容

在检索到的86项研究中，有17项使用dPCR应用于临床人类感兴趣的原生动物（8），蠕虫（8）和节肢动物（1）的寄生虫。在四项研究中分别采用dPCR进行了疟原虫和日本血吸虫的研究。dPCR在恶性疟原虫和恶性疟原虫中以及在日本血吸虫中显示出优于定量实时PCR的明显优势；在隐孢子虫中对粪便抑制剂的稳定性更高。对于所有寄生虫，dPCR都可以进行绝对定量而无需标准曲线。使用了各种dPCR平台。需要考虑一些关键因素：DNA负载，平台选择和反应优化。

含意

由于其灵敏度和定量特性，dPCR有望成为将来在寄生虫检测和定量分析分子技术中吸引人的新方法。通常，它比仅基因组DNA检测有更多的应用，例如混合感染中的定量，基因表达和突变分析。dPCR在疟疾筛查和诊断中应被视为对常规检测和血吸虫病消除计划的补充。dPCR在常规临床实验室中的整合需要标准化的策略和进一步的研究。