Simple Sample Preparation Method for Direct Microbial Identification and Susceptibility Testing From Positive Blood Cultures.
Simple Sample Preparation Method for Direct Microbial Identification and Susceptibility Testing From Positive Blood Cultures.
复制标题
用于直接微生物鉴定和阳性血培养物药敏试验的简单样品制备方法
DOI：
10.3389/fmicb.2018.00481
复制
发表时间：
2018
影响因子：
5.2
通讯作者：
Sun EH
中科院分区：
文献类型：
--
作者：
Pan HW;Li W;Li RG;Li Y;Zhang Y;Sun EH
快速鉴定和测定血液感染患者感染病原体的药敏图谱是选择有效的靶向抗生素进行治疗的关键步骤。然而，在开发同时直接鉴定和确定阳性血培养中细菌的抗生素敏感性的组合方法方面所做的工作很少。在本研究中，我们建立了裂解-离心-洗涤程序，从阳性血培养中制备细菌颗粒，该颗粒可直接用于基质辅助激光解吸/电离飞行时间质谱仪(MALDI-TOF MS)的鉴定和VITEK 2系统的药敏试验。对该方法进行了评估，总共使用了129份临床细菌阳性的血液培养。整个样品制备过程可在15分钟内完成。直接MALDI-TOF MS鉴定革兰氏阴性菌的正确率为96.49%，阳性菌的正确率为97.22%。VITEK-2革兰氏阴性菌药敏试验结果显示，抗菌药物分类符合率为96.89%，小错误率为2.63%，大错误率为0.24%，极大错误率为0.24%。抗菌药物对革兰氏阳性菌的分类符合率为92.81%，小错误率为4.51%，大错误率为1.22%，极大错误率为1.46%。这些结果表明，与传统的依赖培养的实验室方法相比，我们的直接抗生素敏感性分析方法工作得很好。总体而言，这种快速、简便、准确的方法可以方便地对阳性血培养中的细菌进行直接鉴定和药敏试验。
Rapid identification and determination of the antibiotic susceptibility profiles of the infectious agents in patients with bloodstream infections are critical steps in choosing an effective targeted antibiotic for treatment. However, there has been minimal effort focused on developing combined methods for the simultaneous direct identification and antibiotic susceptibility determination of bacteria in positive blood cultures. In this study, we constructed a lysis-centrifugation-wash procedure to prepare a bacterial pellet from positive blood cultures, which can be used directly for identification by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS) and antibiotic susceptibility testing by the Vitek 2 system. The method was evaluated using a total of 129 clinical bacteria-positive blood cultures. The whole sample preparation process could be completed in <15 min. The correct rate of direct MALDI-TOF MS identification was 96.49% for gram-negative bacteria and 97.22% for gram-positive bacteria. Vitek 2 antimicrobial susceptibility testing of gram-negative bacteria showed an agreement rate of antimicrobial categories of 96.89% with a minor error, major error, and very major error rate of 2.63, 0.24, and 0.24%, respectively. Category agreement of antimicrobials against gram-positive bacteria was 92.81%, with a minor error, major error, and very major error rate of 4.51, 1.22, and 1.46%, respectively. These results indicated that our direct antibiotic susceptibility analysis method worked well compared to the conventional culture-dependent laboratory method. Overall, this fast, easy, and accurate method can facilitate the direct identification and antibiotic susceptibility testing of bacteria in positive blood cultures.
登录
查看更多内容
DOI：
10.1016/j.jmii.2017.03.005
发表时间：
2018-10-01
影响因子：
7.4
作者：
Lin, Jung-Fu;Ge, Mao-Cheng;Lu, Jang-Jih
通讯作者：
Lu, Jang-Jih
影响因子：
28.2
作者：
Ascione, T.;Pagliano, P.;Esposito, S.
通讯作者：
Esposito, S.
影响因子：
9.4
作者：
Chen, Jonathan H. K.;Ho, Pak-Leung;Yam, Wing-Cheong
通讯作者：
Yam, Wing-Cheong
DOI：
10.1016/j.diagmicrobio.2012.01.010
发表时间：
2012-05-01
影响因子：
2.9
作者：
Saffert, Ryan T.;Cunningham, Scott A.;Patel, Robin
通讯作者：
Patel, Robin
影响因子：
2.9
作者：
Judd, William R.;Stephens, Dana M.;Kennedy, Charles A.
通讯作者：
Kennedy, Charles A.