Ryba S, Tacner J, Havlickova M, Stopka P
Acta Virol. 2012;56(2):125-8. doi: 10.4149/av_2012_02_125.
Various methods of handling samples of avian influenza prior to detecting influenza viruses can significantly influence both, the detection of the virus and the quantification of viral nucleic acids. The quantity of influenza viral RNA remaining in different collecting buffers and kept at temperatures of -20°C, +4°C or +22°C for various lengths of time, was determined. The quantity of viral RNA remained the same for 120 days at -20°C, but decreased when the samples were stored at either +4°C or +22°C. The quantity of RNA was influenced by the composition of the collecting buffer. The influenza virus sample that is to be used for RNA quantification can be stored at +4°C and freeze and thaw cycles should be avoided during transport. Our results clearly indicate that the quality and quantity of influenza virus nucleic acid depends on the chemical composition of used buffer and also that the samples can be protected from degradation even if they are not stored at ultra-low temperatures. However, repeated thaw and freeze cycles will damage viral RNA even if kept in stabilizing buffers.
influenza virus; degradation; RNA; buffer.
在检测流感病毒之前,处理禽流感样本的各种方法会对病毒检测和病毒核酸定量产生显著影响。测定了在不同收集缓冲液中、于-20°C、+4°C或+22°C温度下保存不同时长后剩余的流感病毒RNA量。在-20°C下,病毒RNA量在120天内保持不变,但当样本保存在+4°C或+22°C时会减少。RNA量受收集缓冲液成分的影响。用于RNA定量的流感病毒样本可保存在+4°C,运输过程中应避免冻融循环。我们的结果清楚地表明,流感病毒核酸的质量和数量取决于所用缓冲液的化学成分,而且即使样本不是保存在超低温下,也可防止其降解。然而,即使保存在稳定缓冲液中,反复的冻融循环也会损害病毒RNA。
流感病毒;降解;RNA;缓冲液