娄静, 刁保卫, 李伟, 崔志刚, 阚飙. 三种核酸染料应用于细菌脉冲场凝胶电泳染色的比较研究[J]. 疾病监测, 2011, 26(5): 388-391.
引用本文: 娄静, 刁保卫, 李伟, 崔志刚, 阚飙. 三种核酸染料应用于细菌脉冲场凝胶电泳染色的比较研究[J]. 疾病监测, 2011, 26(5): 388-391.
LOU Jing, DIAO Bao-wei, LI Wei, CUI Zhi-gang, KAN Biao. Comparative analysis of three types of nucleic acid staining for PFGE[J]. Disease Surveillance, 2011, 26(5): 388-391.
Citation: LOU Jing, DIAO Bao-wei, LI Wei, CUI Zhi-gang, KAN Biao. Comparative analysis of three types of nucleic acid staining for PFGE[J]. Disease Surveillance, 2011, 26(5): 388-391.

三种核酸染料应用于细菌脉冲场凝胶电泳染色的比较研究

Comparative analysis of three types of nucleic acid staining for PFGE

  • 摘要: 目的 比较Gelred、GoldView及溴化乙锭(EB)三种染料在病原菌脉冲场凝胶电泳(PFGE)中的染色效果,选择更为安全和灵敏的核酸染料,并形成标准化操作应用于病原菌分子分型监测网络PulseNet China。 方法 选择霍乱弧菌作为评价菌株,按照病原菌分子分型监测网络(PulseNet)的PFGE标准操作方案进行电泳,采用Gelred、GoldView及EB三种染料分别染色,分析比较实验结果。 结果 Gelred染色的胶块清晰度最好,EB次之,均高于GoldView; Gelred染色的图谱,相邻条带边缘清晰,更容易辨别区分。EB和GoldView染色后必须脱色,使用Gelred染色脱色时间较短,亦可以不经过脱色步骤,但脱色后的电泳条带边缘更加锐利。 结论 Gelred是一种更安全、更灵敏的核酸染料,可以取代EB应用于PFGE。制定了使用Gelred进行PFGE染胶的标准方案,用于PulseNet China网络的PFGE分析。

     

    Abstract: Objective To compare the safety and sensitivity of three dyes, including Gelred, GoldView and Ethidium bromide (EB), used as nucleic acid staining reagents in pulsed-field gel electrophoresis (PFGE) for pathogens, and optimize and standardize the protocol of the molecular typing surveillance network of pathogens-PulseNet China. Methods Vibrio cholera isolates were selected for evaluation purpose. PFGE was carried out according to the standard protocol on molecular typing surveillance network (PulseNet). The agarose gels after electrophoresis were stained by Gelred, GoldView and EB, respectively. The staining effects were observed. Results It is showed that the staining effect of Gelred was best among the three tested dyes. The observed DNA bands in the gels stained by Gelred is most clear, followed by EB and GoldView. The band edge showed by Gelred staining was more clear and distinguishable. It would take time for gels to be destained after EB and GoldView staining, but if Gelred is used, the destaining time could be shortened or be neglected. Even though, it is noticeable that the edge of destained electrophoretic band after Gelred staining would be more clear. Conclusion The results suggested that Gelred as a nucleic acid staining dye is safer and more sensitive for gel viewing in PFGE procedure, and is promising to replace EB to develop PFGE standard protocol in PulseNet China network.

     

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