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环介导恒温扩增技术快速检测短凯伦藻

DOI: 10.3724/SP.J.1145.2012.00482, PP. 482-488

Keywords: 短凯伦藻,环介导恒温扩增,特异性,灵敏性,赤潮微藻,短藻毒素

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Abstract:

短凯伦藻(kareniabrevis)是一种有毒赤潮微藻,所产生的短藻毒素对海洋生物乃至人类都有毒害作用,为加强对短凯伦藻赤潮的监控,建立了稳定的短凯伦藻环介导恒温扩增(lamp)鉴定体系,在此基础上进行了特异性和灵敏性验证.特异性实验结果显示只有短凯伦藻或者含短凯伦藻的模板呈现阳性反应,其他微藻为阴性反应,从而验证了该lamp方法的特异性;同时,对短凯伦藻的基因组dna进行一系列10倍稀释作为敏感度实验的模板,并与常规pcr做了对比,结果表明:短凯伦藻的lamp方法最低检测限度为50pg,敏感度比常规pcr高10倍.lamp产物鉴定不需要常规的胶电泳过程,直接采用肉眼观察的方法,在含有短凯伦藻的阳性反应管中会出现白色混浊,加入sybrògreeni染料呈现绿色,而未含有短凯伦藻的阴性管为澄清,染色后仍为原来的橙色.因此,该方法操作简便、特异性强、灵敏度高而成本低,在赤潮原因种检测监控方面具有良好的应用前景.

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