免疫捕获PCR法快速检测金黄色葡萄球菌

Rapid Detection of Staphylococcus aureus by Immunocaptured PCR

旨在建立一种快速检测金黄色葡萄球菌(Staphylococcus aureus,SA)免疫捕获PCR技术,并探讨其灵敏度和特异性。SA特异性抗体包被PCR管以富集待测样品中目标菌,之后在同一PCR管里直接进行免疫捕获PCR,并和直接PCR比较。免疫捕获PCR法可特异性检测2株SA菌株,而无法检测到其它8种常见食源性致病菌,说明该方法对SA具有良好的特异性;该方法对纯菌液而言,检测灵敏度可达到2.35×102CFU/mL,是直接PCR的100倍;对5种食品模拟带菌检测发现,无需增菌培养,其灵敏度可达到2.35×103-2.35×104CFU/mL,是直接PCR的10-100倍。免疫捕获PCR法集免疫学与分子生物学检测技术于一体,具有高特异性、高灵敏度、检测快速、易于操作、成本低廉等诸多优点,是一种适合基层实验室使用的检测技术。

It was to establish the Immunocapture PCR technique to detect staphylococcus aureus （ SA ） , further investigate its sensitivity and specificity. PCR tubes coated with SA antibodies for enriching target bacteria and directly amplification, then Immunocapture PCR compared with direct-PCR. It found that two strains of SA showed positive results by the method of immumnocapture PCR, whereas other eight foodborne pathogens strains showed negative results. Positive detection of SA was obtainable when the load of organism in the test samples was over 2.35 x 102 CFU/mL, and the sensitivity is 100-fold than direct-PCR. Five kinds of food artificially contaminated without enrichment, the sensitivity can reaeb 2.35 x 103 -2.35 ~ 104 CFU/mL, and the sensitivity is 10-100 fold than direct-PCR. Immunooapture PCR method combined immunology with molecular biology technology, with high specificity, high sensitivity, rapid, easy operation and low cost advantages, could be suitable for primary laboratory.