Departamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional (IPN), Mexico City, Mexico.
Departamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional (IPN), Mexico City, Mexico.
J Virol Methods. 2019 Jan;263:44-49. doi: 10.1016/j.jviromet.2018.10.013. Epub 2018 Oct 19.
Retroviral pseudotypes are broadly used as safe instruments to mimic the structure and surface of highly pathogenic viruses. They have been employed for the discovery of new drugs, as diagnostic tools in vaccine studies, and part of serological assays. Because of their widespread use in research and their potential as tools for quality control, it is important to know their shelf life, stability, and best storage conditions. In this study, we produced pseudotypes carrying the lacZ reporter gene and the hemagglutinin (HA) of avian influenza virus subtypes H5 and H7 to investigate their stability under various storage conditions. We produced pseudotypes with titers of approximately 10 RLU/mL, which decreased to 10-10 RLU/mL after short-term storage at 4 °C (up to 4 weeks). Stability was maintained after long-term storage at -20 °C (up to 12 months), even under storage variations such as freeze-thaw cycles. We conclude that, although the titers decreased by 1 log under the different storage conditions, the remaining titers can be readily applicable in many techniques, such as neutralization assays. These findings show that large quantities of retroviral pseudotypes can be safely stored for short- or long-term use, allowing standardization and reduced variation in assays involving retroviral pseudotypes.
逆转录病毒假型广泛用作模拟高致病性病毒结构和表面的安全工具。它们已被用于新药的发现、疫苗研究中的诊断工具以及血清学检测的一部分。由于它们在研究中的广泛应用及其作为质量控制工具的潜力，了解它们的保质期、稳定性和最佳储存条件非常重要。在这项研究中，我们生产了携带 lacZ 报告基因和禽流感病毒亚型 H5 和 H7 的血凝素 (HA) 的假型，以研究它们在各种储存条件下的稳定性。我们生产的假型滴度约为 10 RLU/mL，在 4°C（长达 4 周）的短期储存后降至 10-10 RLU/mL。在 -20°C（长达 12 个月）的长期储存下保持稳定，即使在储存变化（如冻融循环）下也是如此。我们得出结论，尽管在不同的储存条件下滴度下降了 1 个对数，但剩余的滴度可在许多技术中广泛应用，如中和测定。这些发现表明，大量的逆转录病毒假型可以安全储存用于短期或长期使用，从而实现涉及逆转录病毒假型的测定的标准化和减少变异。
