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氰基藻芯片:一种高通量检测环境蓝藻的新型 DNA 微阵列工具,可在菌株水平上进行。

CyanoStrainChip: A Novel DNA Microarray Tool for High-Throughput Detection of Environmental Cyanobacteria at the Strain Level.

机构信息

Guangdong Magigene Biotechnology Co., Ltd., Shenzhen 518081, PR China.

School of Food and Drug, Shenzhen Polytechnic, Shenzhen 518081, PR China.

出版信息

Environ Sci Technol. 2024 Mar 19;58(11):5024-5034. doi: 10.1021/acs.est.3c11096. Epub 2024 Mar 7.

Abstract

Detecting cyanobacteria in environments is an important concern due to their crucial roles in ecosystems, and they can form blooms with the potential to harm humans and nonhuman entities. However, the most widely used methods for high-throughput detection of environmental cyanobacteria, such as 16S rRNA sequencing, typically provide above-species-level resolution, thereby disregarding intraspecific variation. To address this, we developed a novel DNA microarray tool, termed the CyanoStrainChip, that enables strain-level comprehensive profiling of environmental cyanobacteria. The CyanoStrainChip was designed to target 1277 strains; nearly all major groups of cyanobacteria are included by implementing 43,666 genome-wide, strain-specific probes. It demonstrated strong specificity by mock community experiments. The high correlation (Pearson's > 0.97) between probe fluorescence intensities and the corresponding DNA amounts (ranging from 1-100 ng) indicated excellent quantitative capability. Consistent cyanobacterial profiles of field samples were observed by both the CyanoStrainChip and next-generation sequencing methods. Furthermore, CyanoStrainChip analysis of surface water samples in Lake Chaohu uncovered a high intraspecific variation of abundance change within the genus between different severity levels of cyanobacterial blooms, highlighting two toxic strains that are of critical concern for Lake Chaohu harmful blooms suppression. Overall, these results suggest a potential for CyanoStrainChip as a valuable tool for cyanobacterial ecological research and harmful bloom monitoring to supplement existing techniques.

摘要

由于蓝藻在生态系统中起着至关重要的作用,因此检测环境中的蓝藻是一个重要的关注点。它们可能会形成水华,从而对人类和非人类实体造成危害。然而,目前用于高通量检测环境蓝藻的最广泛方法,如 16S rRNA 测序,通常只能提供高于物种水平的分辨率,从而忽略了种内变异。为了解决这个问题,我们开发了一种新的 DNA 微阵列工具,称为 CyanoStrainChip,它能够对环境蓝藻进行菌株水平的综合分析。CyanoStrainChip 是针对 1277 株菌株设计的;通过实施 43666 个全基因组、菌株特异性探针,几乎涵盖了所有主要的蓝藻群体。通过模拟群落实验,它表现出很强的特异性。探针荧光强度与相应 DNA 量(从 1-100ng 不等)之间的高度相关性(Pearson's > 0.97)表明其具有出色的定量能力。CyanoStrainChip 和下一代测序方法都观察到了现场样本的一致蓝藻图谱。此外,对巢湖地表水样本的 CyanoStrainChip 分析揭示了在不同蓝藻水华严重程度下,属内丰度变化的高度种内变异,突出了两种具有毒性的菌株,这对巢湖有害水华的抑制具有重要意义。总的来说,这些结果表明 CyanoStrainChip 有可能成为蓝藻生态研究和有害水华监测的一种有价值的工具,以补充现有的技术。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6671/10956431/4e8e67dba4d4/es3c11096_0001.jpg

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