混合感染的多种亚型禽流感病毒的纯化与鉴定

Purification and identification of avian influenza viruses causing mixed multi-infection

摘要:

摘要：【目的】研究混合感染的多种亚型禽流感病毒的纯化和鉴定方法。【方法】用鸡胚终点稀释法和鸡胚终点稀释法结合特异性血清中和法分别对2-3种已知亚型禽流感病毒的混合感染样品进行纯化，并对纯化结果用RT-PCR和血凝抑制试验进行鉴定。用建立的方法对214份禽流感病毒阳性样品进行了纯化和鉴定。【结果】用鸡胚终点稀释法对样品稀释、传代6-7次可使病毒达到纯化，但用鸡胚终点稀释法结合特异性血清中和法对样品稀释、传代4-5次即可达到病毒纯化。用RT-PCR和血凝抑制试验两种方法同时鉴定病毒的纯化效果，可明显提高准确性。用本方法从214份样品中纯化出涵盖13种亚型的禽流感病毒233株。【结论】鸡胚终点稀释法及其结合特异性血清中和法均能对禽流感病毒进行纯化，但是鸡胚终点稀释法结合特异性血清中和法更具有针对性，也更有效。另外，对纯化结果的鉴定需采取多种手段。

Abstract:

Abstract: [Objective] To purify and identify avian influenza viruses causing multi-subtype mixed infection. [Methods] By using chicken embryo end-point dilution or combining with specific antiserum neutralizing tests, the mix-infection samples of 2 or 3 subtyping avian influenza viruses were purified, and the results of purification were identified by RT-PCR and hemagglutination inhibition tests. These methods were used to purify and identify 214 positive samples of avian influenza viruses. [Results] The method of chicken embryo end-point dilution was able to purify the viruses from samples by dilution and subculturing six or seven passages. However, combining this method with the specific antiserum neutralizing tests, the samples could be purified after four or five passages. The accuracy of identification could be obviously enhanced by using RT-PCR and hemagglutination inhibition tests simultaneously. Based on these methods, 233 of avian influenza viruses, which constituted 13 subtypes, were purified from 214 samples. [Conclusion] Both chicken embryo end-point dilution method and its combination with specific antiserum neutralizing tests could be used to purify avian influenza viruses of mix-infection, however, the latter may be more focused and valid. In addition, the results of purification should also be evaluated by multiple means.