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首页> 外文期刊>Harmful Algae >Development of a PNA probe for the detection of the toxic dinoflagellate Takayama pulchella
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Development of a PNA probe for the detection of the toxic dinoflagellate Takayama pulchella

机译：开发用于检测有毒的鞭毛鞭毛高山龟的PNA探针

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摘要

A peptide nucleic acid (PNA) probe was developed to detect the toxic dinoflagellate, Takayama pulchella TPXM, using fluorescent in situ hybridization (FISH) combined with epifluorescent microscopy and flow cytometry. The PNA probe was then used to analyze HAB samples from Xiamen Bay. The results indicated that the fluorescein phosphoramidite (FAM)-labeled probe (PNATP28S01) [Flu]-OO ATG CCATCT CAA GA, entered the algal cells easily and bound to the target species specifically. High hybridization efficiency (nearly 100%) was observed. Detection by epifluorescence microscopy and flow cytometry gave comparable results. The fluorescence intensity of the PNA probe hybridized to T. pulchella cells was remarkably higher than that of two DNA probes used in this study and than the autofluorescence of the blank and negative control cells. In addition, the hybridization condition of the PNA probe was easier to control than DNA probes, and when applied to field-collected samples, the PNA probe showed higher binding efficiency to the target species than DNA probes. With the observed high specificity, binding efficiency, and detection signal intensity, the PNA probe will be useful for monitoring harmful algal blooms of T. pulchella.
机译：使用荧光原位杂交（FISH）与落射荧光显微镜和流式细胞术相结合，开发了一种肽核酸（PNA）探针来检测有毒的鞭毛藻高山山梨TPXM。然后将PNA探针用于分析厦门湾的HAB样品。结果表明，荧光素亚磷酰胺（FAM）标记的探针（PNATP28S01）[Flu] -OO ATG CCATCT CAA GA容易进入藻类细胞，并与目标物种特异性结合。观察到高的杂交效率（接近100％）。通过落射荧光显微镜和流式细胞仪检测得出可比的结果。与P.pulchella细胞杂交的PNA探针的荧光强度显着高于本研究中使用的两种DNA探针的荧光强度，并且高于空白和阴性对照细胞的自发荧光。此外，PNA探针的杂交条件比DNA探针更容易控制，当应用于野外采集的样品时，PNA探针显示出比DNA探针更高的与靶标物种的结合效率。凭借观察到的高特异性，结合效率和检测信号强度，PNA探针将可用于监测P.chulchella的有害藻华。

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