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开发一种用于非洲家畜血液病原体物种共感染高通量分析的新型血液生物群落工具。

Development of a novel Haemabiome tool for the high-throughput analysis of haemopathogen species co-infections in African livestock.

作者信息

Yalcindag Erhan, Vasoya Deepali, Hemmink Johanneke D, Karani Benedict, Hernandez Castro Luis Enrique, Callaby Rebecca, Mazeri Stella, Paxton Edith, Connelley Timothy K, Toye Phil, Morrison Liam J, Bronsvoort Barend Mark de C

机构信息

Centre for Tropical Livestock Genetics and Health (CTLGH), Roslin Institute, University of Edinburgh, Edinburgh, United Kingdom.

The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Edinburgh, United Kingdom.

出版信息

Front Vet Sci. 2024 Dec 20;11:1491828. doi: 10.3389/fvets.2024.1491828. eCollection 2024.

Abstract

One of the principal limitations on livestock productivity in sub-Saharan Africa is the constraining effect of infectious diseases, including tick-borne blood pathogens. Currently, diagnostic markers for these pathogens are species or genus specific, making it challenging to implement high-throughput screening methods. The aim of this study was to develop and validate a novel high-throughput diagnostic tool capable of detecting a range of important haemopathogens in livestock. To achieve this, we developed a high-throughput diagnostic tool that can detect all species of , and present in a sample. The approach involves targeting the 16S/18S rDNA region by PCR and subjecting amplicons to deep sequencing, which allows for the identification of species present in a sample, and the exploration of haemopathogen communities. To validate the accuracy of this Next Generation Sequencing method, we compared the amplicon sequencing results with species-specific PCR and reverse line blot (RLB) test data of both control and field samples. The Haemabiome tool demonstrated the successful resolution of positive and negative samples, and highlighted the power of this diagnostic tool in identifying multiplicity of infections. The Haemabiome tool can therefore generate valuable insights regarding the understanding of the true diversity of species composition and the distribution of pathogen communities in field samples.

摘要

撒哈拉以南非洲地区家畜生产力的主要限制因素之一是传染病的制约作用,包括蜱传血液病原体。目前,这些病原体的诊断标志物是物种或属特异性的,这使得实施高通量筛查方法具有挑战性。本研究的目的是开发并验证一种新型高通量诊断工具,能够检测家畜中一系列重要的血液病原体。为实现这一目标,我们开发了一种高通量诊断工具,可检测样本中存在的所有 、 和 物种。该方法包括通过PCR靶向16S/18S rDNA区域,并对扩增子进行深度测序,从而能够识别样本中存在的物种,并探索血液病原体群落。为了验证这种下一代测序方法的准确性,我们将扩增子测序结果与对照样本和现场样本的物种特异性PCR及反向线杂交(RLB)测试数据进行了比较。Haemabiome工具成功分辨了阳性和阴性样本,并突出了这种诊断工具在识别多重感染方面的能力。因此,Haemabiome工具可以为了解野外样本中物种组成的真实多样性和病原体群落分布提供有价值的见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f8e/11695320/5c67bad9edaa/fvets-11-1491828-g001.jpg

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