检测霍乱弧菌的基因芯片的制备
Detection of vibrio cholerae with DNA microchips
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摘要:目的 研制快速、特异、灵敏的检测霍乱弧菌基因芯片。方法 选择霍乱弧菌特异的编码外膜蛋白的ompW基因和毒素ctxA基因, 设计引物和探针, 并制备检测芯片, 通过2次PCR扩增, 制备荧光标记的靶序列, 并与芯片进行杂交, 检测霍乱弧菌和非霍乱弧菌病原体。结果 所有霍乱弧菌菌株在采用单一和多重PCR2种方法制备的荧光标记靶序列与芯片杂交, 均在芯片相应探针处出现阳性信号, 非霍乱弧菌菌株杂交结果均为阴性。结论 霍乱弧菌基因检测芯片的研制, 为霍乱弧菌感染的诊断提供了准确、快速、灵敏的方法, 还可选择更多的基因位点设计探针以提高检测的可靠性, 可以达到高通量、集成化和自动化的要求。Abstract:Objective Rapid specific and sensitive DNA microchips are used for detection of vibrio cholerae.Methods Primers and specific probes were designed from vibrio cholerae gene ompW and ctxA.The fluorescent-labeled PCR products were prepared by incorporation of Cy3-labeled primer during PCR amplification and were hybridized to the microchips for detection of vibrio cholerae strains and non vibrio cholerae pathogens.Results Positive signal were all displayed on the mycrochips in vibrio cholerae strains when used conventional PCR or multiplex PCR, but non vibrio cholerae pathogens failed to yield any signal under comparable conditions.Conclusion DNA microchips provide a more specific, rapid and efficient method for vibrio cholerae detection.It might be very useful for high-throughout, integrated and automated identification and detection of bacterial pathogens by designing more probes.
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