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目的 通过RT PCR及病毒分离等方法 ,鉴定SARS感染性动物模型是否成立 ,并对疫苗效果进行评估。方法 选用 3周岁的健康SPF级恒河猴 8只 ,分为SARS实验感染动物模型组及疫苗组 (Sino 3SARS灭活疫苗2 0 0 30 90 4批肌内注射 ,免疫 2次 )。经鼻吸入Sino 1SARS毒株 ,病毒攻击后 7d动物进行安乐死。自病毒攻击后的第一天起每日连续采集动物咽拭子标本 ,病毒攻击后的第二天、第五天、第七天采集血浆标本[1 ] 。将标本进行病毒分离及RT PCR检测 .结果 模型组动物病毒攻击后第一天至第七天咽拭子RT PCR检测均为阳性。注射疫苗组动物仅在病毒攻击后第一天咽拭子RT PCR检测呈阳性。模型组动物血浆RT PCR检测呈现阳性 ,疫苗组动物血浆RT PCR检测均为阴性。病毒分离结果显示仅在模型动物咽拭子标本中分离到病毒 ,未在疫苗组动物的咽拭子标本分离到病毒 .血浆标本中均未分离到病毒 .结论 RT PCR及病毒分离可以及时有效地检测出病毒在体内的复制情况 ,可作为模型鉴定、疫苗评估及药物筛选的重要手段。
Objective To identify whether the SARS-infected animal model is established by RT-PCR and virus isolation and evaluate the vaccine efficacy. Methods Eight healthy 3-year-old healthy SPF rhesus monkeys were divided into 3 groups: animal model group and vaccine group with SARS infection (2000090904 inactivated vaccine of Sino 3SARS). Sino 1SARS strain was inhaled nasally and animals were euthanized 7 days after virus challenge. Throat swab samples were collected daily from the first day after the virus attack, and plasma samples were collected on the second, fifth and seventh days after virus challenge. The specimens were subjected to virus isolation and RT PCR detection.Results The throat swab RT PCR detection was positive on the first day to the seventh day after animal virus challenge in the model group. The throat swab RT-PCR test was positive in the vaccinated animals only on the first day after the virus challenge. In the model group, plasma RT-PCR showed positive results in plasma and RT-PCR in plasma of the vaccine group was negative. Virus isolation results showed that the virus was isolated from throat swab specimens of model animals only, and no virus was isolated from the throat swab specimens of the vaccine animals.No virus was isolated from the plasma samples.Conclusion RT PCR and virus isolation can be performed in a timely and effective manner Detection of virus replication in the body, can be used as a model identification, vaccine evaluation and drug screening an important means.