Development of a high-throughput minimum inhibitory concentration (HT-MIC) testing workflow.
Development of a high-throughput minimum inhibitory concentration (HT-MIC) testing workflow.
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DOI：
10.3389/fmicb.2023.1079033
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发表时间：
2023
影响因子：
5.2
通讯作者：
中科院分区：
文献类型：
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作者：
最低抑菌浓度（MIC）测定的根源可以追溯到20世纪初。从那时起，该测试经历了修改和改进，以提高其可靠性和准确性。虽然生物调查使用的样本数量不断增加，但复杂的过程和人为错误有时会导致数据质量差，这使得复制科学结论具有挑战性。使用机器可破译的协议自动化手动步骤可以减轻程序上的困难。最初依靠手动移液和人类视觉来确定结果，现代肉汤稀释MIC测试程序已纳入酶标仪以增强样品分析。然而，目前的MIC测试程序无法同时有效地评估大量样品。在这里，我们使用Opentrons OT-2机器人创建了一个概念验证工作流程，以实现高通量MIC测试。我们通过将Python编程用于MIC分配来进一步优化分析，以简化自动化。在该工作流程中，我们对4种不同菌株进行了MIC检测，每种菌株重复检测3次，共分析了1，152个威尔斯孔。将我们的工作流程与传统的平板MIC程序进行比较，我们发现HT-MIC方法的速度快800%，同时具有100%的准确性。我们的高通量MIC工作流程可以适应学术和临床环境，因为它比许多传统方法更快，更有效，更准确。
The roots of the minimum inhibitory concentration (MIC) determination go back to the early 1900s. Since then, the test has undergone modifications and advancements in an effort to increase its dependability and accuracy. Although biological investigations use an ever-increasing number of samples, complicated processes and human error sometimes result in poor data quality, which makes it challenging to replicate scientific conclusions. Automating manual steps using protocols decipherable by machine can ease procedural difficulties. Originally relying on manual pipetting and human vision to determine the results, modern broth dilution MIC testing procedures have incorporated microplate readers to enhance sample analysis. However, current MIC testing procedures are unable to simultaneously evaluate a large number of samples efficiently. Here, we have created a proof-of-concept workflow using the Opentrons OT-2 robot to enable high-throughput MIC testing. We have further optimized the analysis by incorporating Python programming for MIC assignment to streamline the automation. In this workflow, we performed MIC tests on four different strains, three replicates per strain, and analyzed a total of 1,152 wells. Comparing our workflow to a conventional plate MIC procedure, we find that the HT-MIC method is 800% faster while simultaneously boasting a 100% accuracy. Our high-throughput MIC workflow can be adapted in both academic and clinical settings since it is faster, more efficient, and as accurate than many conventional methods.
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DOI：
10.3390/s22093242
发表时间：
2022-04-23
期刊：
Sensors (Basel, Switzerland)
影响因子：
--
作者：
通讯作者：
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影响因子：
4.7
作者：
Tenhaef, Niklas;Stella, Robert;Noack, Stephan
通讯作者：
Noack, Stephan
影响因子：
5.2
作者：
Holland, Mische;Bjanes, Elisabet;Dillon, Nicholas
通讯作者：
Dillon, Nicholas
影响因子：
5.7
作者：
Jessop-Fabre, Mathew M.;Sonnenschein, Nikolaus
通讯作者：
Sonnenschein, Nikolaus
影响因子：
5.2
作者：
Bonev, Boyan;Hooper, James;Parisot, Judicael
通讯作者：
Parisot, Judicael