Cryopreservation of virulent Acinetobacter baumannii to reduce variability of in vivo studies.
Cryopreservation of virulent Acinetobacter baumannii to reduce variability of in vivo studies.
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DOI：
10.1186/s12866-015-0580-8
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发表时间：
2015-11-02
期刊：
影响因子：
4.2
通讯作者：
Spellberg B
中科院分区：
文献类型：
--
作者：
Nielsen TB;Bruhn KW;Pantapalangkoor P;Junus JL;Spellberg B
微生物检测需要准确和可重复性的细菌接种物制备。不同的人在不同的日期准备的接种剂在不同的实验中可能会有很大的差异。这种差异对于革兰氏阴性细菌感染尤其有问题，对于这种感染，阈值效应可能会导致宿主结果的显著差异，而接种物的差异很小。我们比较了传统方法和使用冷冻原料制备小鼠感染鲍曼不动杆菌接种物的准确性。标准接种剂制备导致实际接种量与目标接种量相比有很大差异，也与类似接种量的体内结果有很大差异。细菌的超低温保存对细菌的生长没有明显的影响。此外，从冷冻菌种制备多重感染接种物显著提高了所获得的接种物的准确性，并导致感染的体内结果更具重复性。冷冻菌种减少了与接种制剂相关的试验间变异性，没有表现出显著的生长能力损失，并保持了毒力，增加了感染的可靠性。当感染鲍曼不动杆菌时，冷冻库存需要相当少的时间来准备和提高体内实验结果的重现性。
Microbiological assays require accurate and reproducible preparation of bacterial inocula. Inocula prepared on different days by different individuals can vary significantly from experiment to experiment. This variance is particularly problematic for Gram-negative bacterial infections, for which threshold effects can result in marked variations in host outcome with minor differences in inocula. We compared the accuracy of traditional methods versus using frozen stocks for preparing Acinetobacter baumannii inocula for infection in mice. Standard inoculum preparation resulted in substantial variability, both with respect to the actual inocula achieved compared to the targeted inocula, and with respect to the in vivo outcome resulting from similar inocula. Cryopreservation of the bacteria resulted in no significant decrement in growth of the bacteria. Furthermore, preparation of multiple infectious inocula from a frozen stock significantly improved the accuracy of the achieved inocula, and resulted in more reproducible in vivo outcomes from infection. Frozen stocks reduced inter-experiment variability associated with inoculum preparation, displayed no significant loss of growth capacity, and maintained virulence, increasing the reliability of infection. Frozen stocks require considerably less time to prepare and enhance reproducibility of in vivo experimental results when infecting with A. baumannii.