国境口岸金黄色葡萄球菌的NASBA快速检测

Rapid detection of staphylococcus aureus using NASBA assay at frontier port

目的采用NASBA技术,建立国境口岸金黄色葡萄球菌的快速检测方法。方法根据金黄色葡萄球菌核酸酶nuc1基因设计引物,进行NASBA检测,并采用人工添加菌株污染样品进行验证。结果 NASBA技术检测金黄色葡萄球菌特异性强,能够从污染样品中扩增出203bp的特异性片段,试验中未出现假阳性和假阴性结果,灵敏度高,检出限达到4pg/μL总RNA;且检测周期短,NASBA反应在42℃90min即可进行有效扩增,24h即可完成样品中目标菌的筛选。结论因NASBA技术具有特异性强,灵敏度高,检测周期短等优点,不仅适用于国境口岸现场快速检测金黄色葡萄球菌,而且随着该技术的不断发展,未来会有更为广泛的应用。

Objective By using a nucleic acid sequence -based amplification （NASBA） assay, the rapid detec- tion method of Staphylococcus aureus is established at frontier port. Methods A nucleic acid sequence -based amplification （NASBA） for detection of Staphylococcus aureus is carried out according to primers specific to nucl gene, and artificially contaminated samples are used to verify the method. Results The assay could specifically amplify a 203bp fragment from artificially contaminated samples, while neither false positive nor false negative re- sults happened ; the detecting limit of NASBA is 4pg/μl nucleic acid in our test. The assay could amplify effectively on condition of 42℃ in 90min. The detection of Staphylococcus aureus could be completed within 24 hours. Conclusion According to the advantage of specificity, sensitivity and short detection period, the NASBA assay is not only applicable to rapid detection of Staphylococcus aureus at frontier port, and with the development of the technology, the NASBA assay will have more extensive application in the future.