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Sowmya Nagaraj Shylaja Ramlal Joseph Kingston Harsh Vardhan Batra 《Journal of microbiology, immunology, and infection》2018,51(2):191-198
Background/Purpose
Among DNA-based techniques, polymerase chain reaction (PCR) is the most widely accepted molecular tool for the detection of pathogens. However, the technique involves several reagents and multiple pipetting steps that often lead to error-prone results. Additionally, the reagents entail a cold-chain facility to maintain their stability during storage and transportation. The main aim of the present study was to simplify the utility of a pre-optimized multiplex PCR format that was developed to detect toxigenic strains of Staphylococcus aureus by providing stable, pre-mixed, and ready-to-use master mix in a lyophilized formulation.Methods
Master mix containing all reagents except the template was lyophilized in the presence of an excipient lyoprotectant to achieve long-term stability without altering the sensitivity, specificity and PCR performance. Bromophenol blue was also included in the master mix to reduce the risk of external contamination during gel loading. The stability of lyophilized master mix was analyzed at different temperatures. The PCR performance was also examined after exposure of master mix to notable temperature fluctuations during transportation.Results
The shelf-life of lyophilized master mix was estimated to be 1.5 months at ambient temperature and 6 months at 4°C. Stability was unaffected by temperature fluctuations during transportation even in cold-chain-free conditions, thus reducing the cost required for cold storage.Conclusion
The sensitive, cost-effective, ready-to-use, and ambient temperature stable formulation could be implemented as a detection tool in food analysis and diagnostic laboratories and hospitals and for on-field application outside the laboratories, as well as for detection of toxigenic strains of S. aureus. 相似文献2.
目的：找到一种冻干赋形剂，能有效用于蛇毒中抗肿瘤活性物质ACTX-6的保护作用。方法：采用正交实验设计原理和方法，以海藻糖、丙氨酸、NaCl作为冻干赋形剂的组成，筛选了它们的配比，考察样品冻干后的电泳区带和细胞毒性。结果：所找到的赋型剂的配方不仅可以使ACTX-6在冻干后电泳区带和细胞毒性都不改变，并且能够明显改善样品蛋白对动物的毒性，腹腔注射的LD50在加了保护剂后得到提高，静脉给药时局部组织坏死现象得到改善。 相似文献
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目的 为研究5-氟尿嘧啶壳聚糖纳米粒冻干粉的制备工艺,提高5-氟尿嘧啶壳聚糖纳米粒的稳定性.方法 首先制备5-氟尿嘧啶壳聚糖纳米粒,并以外观和再分散性为指标,进行单因素考察并利用正交实验优化工艺.结果 5-氟尿嘧啶壳聚糖纳米粒冻干粉的最佳制备工艺为预冻时间24 h、冻干保护剂为甘露醇、用量为80 mg、浓度为10％.冻干前后包封率差异无统计学意义(P＞0.05),冻干后的粒径和冻干前相比有一定增大.结论 5-氟尿嘧啶壳聚糖纳米粒冻干粉有望成为新剂型. 相似文献
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目的 为研究5-氟尿嘧啶壳聚糖纳米粒冻干粉的制备工艺,提高5-氟尿嘧啶壳聚糖纳米粒的稳定性。方法 首先制备5-氟尿嘧啶壳聚糖纳米粒,并以外观和再分散性为指标,进行单因素考察并利用正交实验优化工艺。结果 5-氟尿嘧啶壳聚糖纳米粒冻干粉的最佳制备工艺为预冻时间24 h、冻干保护剂为甘露醇、用量为80 mg、浓度为10%。冻干前后包封率差异无统计学意义（P>0.05）,冻干后的粒径和冻干前相比有一定增大。结论 5-氟尿嘧啶壳聚糖纳米粒冻干粉有望成为新剂型。 相似文献
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