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11种(株)食源性细菌基因芯片检测方法的建立

, PP. 123-128

Keywords: 食源性细菌,基因芯片,多重PCR,特异性基因,检测

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Abstract:

建立多重PCR反应结合基因芯片技术的11种(株)食源性致病菌检测方法。以金黄色葡萄球菌、志贺菌、霍乱弧菌、单增李斯特菌等11种(株)食源性细菌的特异性基因为靶基因,利用Premier5.0和Oligo6.0软件设计引物和探针,BLAST比对分析特异性。寡核苷酸探针点制醛基玻片制备基因芯片,13对引物分为3组扩增,PCR产物混合后与基因芯片杂交检测。鼠伤寒沙门菌、产气荚膜梭菌、奇异变形杆菌等11种(株)细菌使用该方法检测均能准确鉴定,基因组DNA灵敏度为10--100pg,6组模拟DNA混合样本芯片检测结果与预期一致,18株沙门菌临床分离株检测均为阳性。从样本PCR扩增开始至检测完成,整个操作时间不超过3h。该方法能够对11种(株)食源性致病菌快速准确鉴定,可以满足部分食源性致病菌通量检测的要求,具有较好的临床应用前景。

References

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