摘要
运用表面增强拉曼散射(Surface enhanced Raman scattering,SERS)方法,实现两种大肠埃希菌和两种金黄色葡萄球菌间的高效检测与鉴别。最终为临床上细菌的鉴别与分类提供一种快速有效的鉴别方法。利用微波法制备了适合细菌SERS检测的纳米银溶胶,并以耐甲氧西林金黄色葡萄球菌(Methicillin resistant Staphylococcus aurens,MRSA)为检测样本,优化了银溶胶与菌液的体积比、作用时间,得到最佳测试条件为银溶胶与菌液(麦氏浊度为0.5)体积比为1?2,作用时间为1 h,将其用于两种大肠埃希菌JM109、DH5α和两种金黄色葡萄球菌ATCC25923、MRSA的SERS检测,并对相应特征峰进行归属。分别选择了30例大肠埃希菌JM109、DH5α和30例金黄色葡萄球菌ATCC25923和MRSA作为样本训练集,6例大肠埃希菌JM109、DH5α和6例ATCC25923、MRSA为测试集,基于主成分分析法(PCA)联合线性判别法(LDA)模型,JM109、DH5α训练集分类正确率为93.33%,ATCC25923、MRSA训练集分类正确率为76.67%,盲测集分类正确率分别为91.67%、75%。结果表明,制备的银纳米溶胶作为增强介质测得的不同细菌的SERS图谱使用该判别模型时能较好地区分不同种类的细菌,而对于同种细菌的普通菌和耐药菌的鉴别效果相对较差。本研究为临床上细菌的快速准确鉴别与分类提供参考。
High efficiency detection and identification of two E. coli strains and two Staphylococcus aureus strains was accomplished using surface enhanced Raman scattering (SERS) method. Finally, it provided a rapid and effective identification method for the identification and classification of clinical bacteria. For detecting bacteria SERS nano silver sol was prepared by microwave method, and using MRSA (Methicillin resistant Staphylococcus aureus ) as test samples to optimize the silver sol and the broth volume ratio, reaction time, the optimal testing conditions were obtained as follows: silver colloids and bacteria (Maxwell turbidity 0.5) volume ratio was 1 2, the reaction time one hour, applied in SERS detection of two E. coli JM109, DH5α and two S. aureus ATCC25923, MRSA, and the belonging to the corresponding peaks. The experiments were selected 30 cases of E. coli JM109, DH5α, and 30 cases of S. aureus ATCC25923 and MRSA as the training set, 6 cases of E. coli JM109, DH5α and 6 cases of S. aureus ATCC25923 and MRSA as the testing set, based on principal component analysis (PCA) combined with linear discriminant analysis (LDA) model. The results showed that the correct classification rate of JM109, DH5α training set was at 93.33%, ATCC25923, the correct classification rate of MRSA training set was at 76.67%, the correct classification rate of spanless set were respectively at 91.67%, 75%. The nano silver sol as the enhanced medium showed different bacterial SERS atlases. When using this discriminant model, it was better to distinguish different kinds of bacteria, but for the same bacterial species, the identification effect of common bacteria and drug-resistant bacteria was relatively poor. This study had important guiding significance for the rapid and accurate identification and classification of bacteria in clinic.
作者
游华建
鲁增辉
石萍
张德利
陈仕江
YOU Hua-jian;LU Zeng-hui;SHI Ping;ZHANG De-li;CHEN Shi-jiang(Chongqing Acad. of Chinese Materia Medica, Chongqing 400065)
出处
《微生物学杂志》
CAS
CSCD
2019年第2期76-82,共7页
Journal of Microbiology
基金
重庆市社会事业与民生保健科技创新专项(KA-17-3)