Abstract：
Cronobacter species are potential pathogens that can contaminate powdered infant formula. C. sakazakii and C. malonaticus are the most common species of Cronobacter associated with infections. This study mined new molecular targets for the detection of C. sakazakii and C. malonaticus by using comparative genome approaches. Specific target genes mngB and ompR were obtained and used to detect C. sakazakii and C. malonaticus, respectively. A novel detection method, termed ladder-shape melting temperature isothermal amplification (LMTIA), was developed and evaluated. The detection limit for pure C. sakazakii DNA was 1 pg per reaction and 1 fg per reaction for C. malonaticus. The C. sakazakii, C. malonaticus, and the reference stains were all correctly identified. The amplicons can be successfully visualized and identified by naked eyes when hydroxy naphthol blue dye (HNB dye) was used in the reaction. Therefore, the LMTIA assays developed in this study showed potential application for microorganism identification and detection.
摘要：
Cronobacter物种是可能污染婴儿配方奶粉的潜在病原体。Sakazakii和malonaticus是与感染相关的Cronobacter的最常见物种。本研究通过使用比较基因组方法挖掘了新的分子靶标，用于检测sakazakii和malonaticus。获得了特定的靶基因mngB和ompR，并用于检测sakazakii和malonaticus，分别。一种新颖的检测方法，称为梯形解链温度等温扩增(LMTIA)，进行了开发和评估。纯的SakazakiiDNA的检测极限为每个反应1μg,而对于马洛氏梭菌,每个反应1μg。theC.Sakazaki,C.malonaticus,和参考染色都被正确识别。当在反应中使用羟基萘酚蓝染料(HNB染料)时,扩增子可以通过肉眼成功地可视化和鉴定。因此,本研究中开发的LMTIA测定法显示了在微生物鉴定和检测中的潜在应用。
