%0 Journal Article
%T 金黄色葡萄球菌DPO-PCR快速检测方法的建立与应用
%A 李丹丹
%A 徐义刚
%A 邱索平
%A 王昱
%A 高会江
%A 高慎阳
%J 食品与生物技术学报
%D 2016
%R 10.3969/j.issn.1673-1689.2016.04.013
%X 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。</br>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
%K 金黄色葡萄球菌 Sa442基因 DPO-PCR&lt
%K /br&gt
%K Staphylococcus aureus
%K Sa442 gene
%K DPO-PCR
%U http://spyswjs.cnjournals.com/spyswjs/ch/reader/view_abstract.aspx?file_no=201604013&flag=1