Lang Qiaoli, Huang Nan, Li Liping, Ge Liangpeng, Yang Xi
Chongqing Academy of Animal Sciences, Chongqing 402460, China.
Sheng Wu Gong Cheng Xue Bao. 2022 May 25;38(5):1994-2002. doi: 10.13345/j.cjb.210624.
Avicel is made of a mixture of microcrystalline cellulose (MCC) and carboxymethyl cellulose (CMC), and used for virus plaque assay. The avicel in common use is produced by FMC Biopolymer. Due to the relatively fixed proportion of MCC and CMC, avicel in common use is not suitable for plaque determination experiment of all types of viruses. In this study, we evaluated the effect of avicel made of different proportions of MCC and CMC on virus plaque assay, and developed an improved avicel virus plaque assay featured with simple and convenient operation, good practicability and high stability. To generate avicel overlays with different proportions of MCC and CMC, twelve different 2×avicel solutions were prepared. Their overall viscosity and bottom viscosity were measured to evaluate the ease of operation. The results showed that most of the 2×avicel solutions (except the 4.8% MCC+1.4% CMC and 4.8% MCC+1.0% CMC group) were easy to absorb and prepare nutrient overlap than 2×CMC solution. In order to find the best scheme to detect the titer of porcine epidemic diarrhea virus (PEDV), these avicel overlay solutions with different proportion of MCC and CMC were used as a replacement in the standard plaque assay. By comparing the size, clarity, stability and titer accuracy of virus plaque, we identified that 0.6% MCC and 0.7% CMC was the most preferable composition of avicel overlay for PEDV plaque assay. In conclusion, we developed an improved virus plaque assay based on avicel, which may facilitate the research of virus etiology, antiviral drugs and vaccines.
微晶纤维素（MCC）和羧甲基纤维素（CMC）的混合物制成了微晶纤维素，用于病毒蚀斑测定。常用的微晶纤维素由FMC生物聚合物生产。由于MCC和CMC的比例相对固定，常用的微晶纤维素不适用于所有类型病毒的蚀斑测定实验。在本研究中，我们评估了由不同比例的MCC和CMC制成的微晶纤维素对病毒蚀斑测定的影响，并开发了一种改进的微晶纤维素病毒蚀斑测定方法，其特点是操作简单方便、实用性好且稳定性高。为了制备具有不同比例MCC和CMC的微晶纤维素覆盖物，制备了十二种不同的2×微晶纤维素溶液。测量它们的总体粘度和底部粘度以评估操作的难易程度。结果表明，大多数2×微晶纤维素溶液（除4.8%MCC + 1.4%CMC和4.8%MCC + 1.0%CMC组外）比2×CMC溶液更容易吸收并制备营养覆盖物。为了找到检测猪流行性腹泻病毒（PEDV）滴度的最佳方案，将这些具有不同比例MCC和CMC的微晶纤维素覆盖溶液用作标准蚀斑测定的替代品。通过比较病毒蚀斑的大小、清晰度、稳定性和滴度准确性，我们确定0.6%MCC和0.7%CMC是PEDV蚀斑测定中微晶纤维素覆盖物的最优选组成。总之，我们开发了一种基于微晶纤维素的改进病毒蚀斑测定方法，这可能有助于病毒病因学、抗病毒药物和疫苗的研究。
