Department of Infectious -Tropical Diseases and Microbiology, IRCCS Sacro Cuore Don Calabria Hospital, Negrar, Verona, Italy.
Department of Infectious -Tropical Diseases and Microbiology, IRCCS Sacro Cuore Don Calabria Hospital, Negrar, Verona, Italy.
Clin Microbiol Infect. 2019 Dec;25(12):1510-1516. doi: 10.1016/j.cmi.2019.06.009. Epub 2019 Jun 18.
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.
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.
Peer-reviewed literature pertinent to this review based on PubMed, Cochrane and Embase databases as well as laboratory experience of authors.
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.
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.
寄生虫感染是导致全球疾病负担的重要原因之一，这是由于国际旅行和移民的结果。在寄生虫病流行地区，为了支持寄生虫病的控制和消除计划以及在非寄生虫病流行地区快速发现病例，需要更准确的诊断工具。数字 PCR（dPCR）是一种强大的技术，最近在寄生虫学中得到了应用。
本综述首次概述了 dPCR 作为一种新型技术应用于寄生虫感染的检测，并强调了该检测方法的最相关潜在益处。
基于 PubMed、Cochrane 和 Embase 数据库以及作者的实验室经验，检索到与本综述相关的同行评议文献。
在检索到的 86 项研究中，有 17 项研究使用 dPCR 应用于属于原生动物（8 项）、蠕虫（8 项）和节肢动物（1 项）的临床人类寄生虫。dPCR 分别在 4 项研究中应用于疟原虫和日本血吸虫。dPCR 在疟原虫和 spp.中比定量实时 PCR 具有更高的灵敏度，在日本血吸虫中具有更高的灵敏度，并且在粪便抑制剂中具有更高的稳定性；在隐孢子虫中具有更高的稳定性。对于所有寄生虫，dPCR 允许进行绝对定量，而无需标准曲线。各种 dPCR 平台都得到了应用。一些关键因素需要考虑：DNA 负荷、平台选择和反应优化。
由于其灵敏度和定量特性，dPCR 可能成为未来寄生虫检测和定量分析的分子技术中一种有吸引力的新方法。一般来说，它的应用不仅仅局限于基因组 DNA 检测，例如在混合感染、基因表达和突变分析中的定量。dPCR 应在疟疾筛查和诊断中作为常规检测的补充，并在血吸虫病消除计划中使用。需要制定标准化策略并进一步研究，以便将 dPCR 纳入常规临床实验室。
