金黄色葡萄球菌DPO-PCR快速检测方法的建立与应用

为建立检测金黄色葡萄球菌（SA）的PCR方法，以SA Sa442基因为靶基因设计了一对双启动寡核苷酸引物（DPO），建立了SA的DPO-PCR 快速检测方法。结果显示，在45～65 ℃退火温度范围内均能高效地扩增出目的基因，表明DPO引物对退火温度不敏感，该方法的灵敏度为2.27×102 cfu/mL，同时DPO 引物特异性强，与其他菌株非特异性扩增反应。利用该方法和行标法分别对采集的175份样品进行检测，均共计检出13份SA阳性样品，两者符合率为100%。作者建立的DPO-PCR 方法设计简单、特异性强，具有良好的实用性，为快速准确检测SA提供新的检测手段。
A dual-priming oligonucleotide (DPO)-based PCR was developed for the rapid detection of Staphylococcus aureus (SA) using Sa442 of SA as a target gene. DPO primers were able to efficiently amplify the target gene in a temperature range from 45 ℃ to 65 ℃，indicating that the DPO-PCR method was not sensitive to the annealing temperature. The DPO-PCR sensitivity was 2.27×102 cfu/mL. Because of a high specificity of DPO-PCR due to the special structures of DPO primers，no non-specific amplifications were produced in reaction. In addition，13 positive samples for SA were detected from 175 clinical samples by the DPO-PCR method，which was in accordance with the result by the SN/T 1870-2007 standard detection protocol. Therefore，the DPO-PCR could be used as a sensitive，rapid and simple tool