Wachiralurpan Sirirat, Sriyapai Thayat, Areekit Supatra, Sriyapai Pichapak, Augkarawaritsawong Suphitcha, Santiwatanakul Somchai, Chansiri Kosum
Department of Biochemistry, Faculty of Medicine, Srinakharinwirot University, Bangkok, Thailand.
Faculty of Environmental Culture and Ecotourism, Srinakharinwirot University, Bangkok, Thailand.
Front Chem. 2018 Apr 3;6:90. doi: 10.3389/fchem.2018.00090. eCollection 2018.
is a major foodborne pathogen of global health concern. Herein, the rapid diagnosis of has been achieved using loop-mediated isothermal amplification (LAMP) based on the phosphatidylcholine-phospholipase C gene (). Colorimetric detection was then performed through the formation of DNA concatemers and a gold nanoparticle/DNA probe complex (GNP/DNA probe). The overall detection process was accomplished within approximately 1 h with no need for complicated equipment. The limits of detection for in the forms of purified genomic DNA and pure culture were 800 fg and 2.82 CFU mL, respectively. No cross reactions were observed from closely related bacteria species. The LAMP-GNP/DNA probe assay was applied to the detection of 200 raw chicken meat samples and compared to routine standard methods. The data revealed that the specificity, sensitivity, and accuracy were 100, 90.20, and 97.50%, respectively. The present assay was 100% in conformity with LAMP-agarose gel electrophoresis assay. Five samples that were negative by both assays appeared to have the pathogen at below the level of detection. The assay can be applied as a rapid direct screening method for .
是一种引起全球健康关注的主要食源性病原体。在此,基于磷脂酰胆碱 - 磷脂酶C基因(),利用环介导等温扩增(LAMP)实现了对的快速诊断。然后通过DNA串联体和金纳米颗粒/DNA探针复合物(GNP/DNA探针)的形成进行比色检测。整个检测过程在约1小时内完成,无需复杂设备。纯化基因组DNA和纯培养物形式的的检测限分别为800 fg和2.82 CFU/mL。未观察到与密切相关细菌物种的交叉反应。LAMP-GNP/DNA探针检测法应用于200份生鸡肉样品的检测,并与常规标准方法进行比较。数据显示,特异性、灵敏度和准确度分别为100%、90.20%和97.50%。本检测法与LAMP-琼脂糖凝胶电泳检测法100%相符。两种检测法均为阴性的5个样品似乎在检测水平以下存在病原体。该检测法可作为对的快速直接筛查方法。