免疫磁珠捕获PCR快速检测单核细胞增生李斯特氏菌

Immunomagnetic Capture PCR for Rapid Detection of Listeria monocytogenes

利用免疫磁珠富集单核细胞增生李斯特氏菌,采用聚合酶链式反应(polymerase chain reaction,PCR)扩增进行快速检测。所制备的免疫磁珠选取在37℃条件下均匀振荡1 h为最佳包被条件,1 mg磁珠偶联抗体的最佳量为100μg/mg,制备的免疫磁珠捕获率为45%。所建立的免疫磁珠捕获-PCR技术在样品细菌浓度达到104 CFU/m L即可被检出,其灵敏度是直接PCR检测限(105 CFU/m L)的10倍,可为病原菌的富集和快速检测提供新方法。

This experiment was conducted using immunomagnetic enrichment followed by amplification with polymerase chain reaction(PCR) for rapid detection of Listeria monocytogenes. The best coating conditions were achieved by shaking the prepared immunomagnetic beads at 37 ℃ for 1 h, and the amount of antibody conjugated to the immunomagnetic beads was 100 μg/mg, leading to a capture rate of 45%. The immunomagnetic capture PCR assay was able to detect as low as 104 CFU/m L Listeria monocytogenes with 10 times higher sensitivity than direct PCR showing a limit of detection of 105 CFU/m L. This PCR assay can provide a new method for the enrichment and rapid detection of pathogenic bacteria.