目的 :建立敏感、特异的西部马脑炎 (westernequineencephalitis,WEE)病毒RT_PCR检测方法。方法 :首先采用逆转录法将病毒基因组RNA逆转录为cDNA ,然后以此cDNA为模板 ,进行PCR扩增。并对PCR扩增过程中的模板量、循环数、Mg2 + 浓度、退火温度和延伸时间等条件进行优化 ,以提高检测的特异性和敏感性。结果 :采用经优化的条件进行PCR扩增获得了约 35 0bp的单一DNA片段 ,其大小与预期的相一致 ,且可自 0 .1TCID50 的病毒液中检测出WEE病毒基因组序列。结论 :所建立的RT_PCR方法可自病毒感染的小鼠脑组织和传代细胞中检测WEE病毒的基因组RNA。 Objective: To establish a sensitive and specific Western blot detection of westernesephalitis (WEE) virus RT_PCR. Methods: Firstly, the viral genomic RNA was reverse transcribed into cDNA by reverse transcription method, then the cDNA was amplified by PCR. The conditions such as the amount of template, the number of cycles, the concentration of Mg2 +, the annealing temperature and the extension time during PCR amplification were optimized to improve the specificity and sensitivity of the detection. Results: A single DNA fragment of about 350 bp was obtained by PCR amplification under the optimal conditions. The size of the single DNA fragment was in accordance with the expectation. WEE virus genomic sequence was detected from 0.1 TCID50 virus solution. CONCLUSION: The established RT_PCR method can detect WEE virus genomic RNA from virus-infected mouse brain tissue and passage cells.