Rotem Shahar, Shifman Ohad, Aloni-Grinstein Ronit
The Department of Biochemistry and Molecular Genetics, The Israel Institute for Biological Research, Ness-Ziona 74100, Israel.
Antibiotics (Basel). 2022 Apr 26;11(5):580. doi: 10.3390/antibiotics11050580.
There is an urgent need for rapid antibiotic susceptibility tests to improve clinical treatment and to support antibiotic stewardship, especially concerning the emergence of multi-drug-resistant bacteria. Nowadays this need is even more profound due to progress in synthetic biology procedures that may facilitate the malicious preparation of engineered antibiotic-resistant pathogens. We recently described a novel, rapid, simple, specific, and sensitive method named a Micro-Agar-PCR-test (MAPt) and showed its performance on clinical as well as environmental samples. The method does not require any isolation or purification steps and is applicable to a wide range of bacterial concentrations, thus allowing a short time to respond within a bioterror event (5-7 h for , 10-12 h for , and 16 h for ). Ready-to-use reagents for this assay may add a level of preparedness. We examined the option of freezing pre-prepared MAPt agar plates and thawing them upon need. Our results show that adequate minimal inhibitory concentration (MIC) values are obtained with the use of thawed 6- and 12-month frozen agar plates. The ability to store MAPt micro-agar plates at -70 °C for a year, together with all other reagents required for MAPt, holds a great advantage for bioterror preparedness.
迫切需要快速的抗生素敏感性测试,以改善临床治疗并支持抗生素管理,尤其是考虑到多重耐药细菌的出现。如今,由于合成生物学程序的进展可能会便利恶意制备工程化的抗生素抗性病原体,这种需求变得更加迫切。我们最近描述了一种名为微琼脂PCR测试(MAPt)的新颖、快速、简单、特异且灵敏的方法,并展示了其在临床以及环境样本上的性能。该方法不需要任何分离或纯化步骤,适用于广泛的细菌浓度范围,从而能够在生物恐怖事件中短时间内做出响应(对于[具体细菌浓度1]为5 - 7小时,对于[具体细菌浓度2]为10 - 12小时,对于[具体细菌浓度3]为16小时)。该检测的即用型试剂可能会提高准备水平。我们研究了冷冻预先制备的MAPt琼脂平板并在需要时解冻的选项。我们的结果表明,使用解冻后的6个月和12个月冷冻琼脂平板可获得足够的最低抑菌浓度(MIC)值。将MAPt微琼脂平板在 - 70°C下储存一年的能力,连同MAPt所需的所有其他试剂,对生物恐怖防范具有很大优势。