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为研究H1亚型猪流感病毒分离株A/Swine/Guangdong/SS1/2012(H1N1,SS1)和A/Swine/Guangdong/YJ28/2014(H1N2,YJ28)的致病力,本研究对2株病毒基因组进行分析,并将2株病毒分别通过滴鼻途径感染仔猪后观察临床症状,在攻毒后4 d与7 d迫杀实验猪采集脏器样品,检测脏器病毒滴度,进行肺病理切片观察和免疫组化实验,比较2株猪流感病毒对猪的致病能力和排毒情况。遗传进化结果显示,SS1分离株8个节段均属于欧亚类禽分支,YJ28为欧亚类禽、pdm/09和人季节性流感的新型重组病毒。动物实验结果显示,两株猪流感病毒均能够在猪只上复制,临床症状均不明显。鼻拭子排毒检测结果显示,YJ28组猪排毒仅持续至感染后第6 d,但SS1组猪在整个实验周期均能检测到排毒。实验猪脏器病毒检测和病理切片结果显示,SS1组相比于YJ28组脏器病毒滴度更高,且造成更严重的肺部病理损伤。本实验分离株SS1相比YJ28在PB2基因出现D~(701)N突变,这或许是SS1比YJ28具有更强致病能力和更长排毒周期的原因。本研究结果为H1N1和H1N2猪流感病毒对猪的致病能力和排毒情况提供实验依据。
In order to study the virulence of A / Swine / Guangdong / SS1 / 2012 (H1N1, SS1) and A / Swine / Guangdong / YJ28 / 2014 (H1N2, YJ28) The genomes were analyzed and the clinical symptoms were observed after inoculating the two strains of virus into the piglets via intranasal route. The experimental pigs were killed 4 days and 7 days after the challenge, respectively. The organ samples were collected for detection of organ virus titer and lung biopsy Observation and immunohistochemistry experiments to compare the two strains of swine influenza virus pathogenicity and detoxification. The results of the genetic evolution showed that all the 8 isolates belonged to the Eurasian poultry branch, and YJ28 was a new recombinant virus of the Eurasian poultry, pdm / 09 and human seasonal flu. Animal experiments show that both swine influenza viruses can replicate in pigs, clinical symptoms are not obvious. Nasal swab detoxification test results showed that detoxification of pigs in YJ28 only lasted 6 days after infection, but swine detoxification was detected in SS1 pigs throughout the experimental period. Experimental pig organ virus detection and biopsy results showed that SS1 group compared to YJ28 group of organ virus titer higher, and cause more serious pathological lung injury. In our study, the mutation of D ~ (701) N occurred in PB2 gene of SS1 compared with that of YJ28, which may be the reason why SS1 has stronger virulence and longer detoxification cycle than YJ28. The results of this study provide experimental evidence for the pathogenicity and detoxification of swine H1N1 and H1N2 swine influenza virus.