三株H1N1亚型猪流感病毒M基因序列测定与进化分析

正猪流感,是集约化养猪场普遍存在且难以根除的猪呼吸道疾病之一,也是世界上最常见的猪传染病之一,目前有10种血清亚型。H1N1亚型SIV在中国日趋复杂和多变,与其他亚型流感病毒密切相关,并在其他谱系之间发生多重重组,其中基质蛋白位于流感病毒第7段,含有1 027个核苷酸,具有M1和M2两个相互重叠的编码区。本试验通过对三株H1N1亚型SIV的M基因进行鉴定分析,为进一步防控猪流感病     

为猪“平反”——甲型H1N1流感防治须知

猪肉卖出白菜价,放在两年前,这是谁都不敢想象的事情。然而受“猪流感”（甲型H1N1流感）事件的影响,市场上猪肉的零售量出现持续下滑,不少养殖大户深受其害。“猪流感”是一种具有高度传染性的猪的急性呼吸道疾病,病毒最常见的是H1N1亚型,但是也存在其他的亚型（如H1N2,H3N1,H3N2）,此次流感并非新病毒,是我们常说的甲1型流感病毒,与1918年在欧洲曾经流行被称为“西班牙流感”相似。专家指出,在人际传播时,打喷嚏、咳嗽和物理接触都有可能导致新型流感病毒在人群间传播,但人不会因吃猪肉或猪产品感染“猪流感”。猪肉加热至71℃,就能杀死“猪流感”病毒,因此我们必须为猪“平反”,改“猪流感”为“甲型H1N1流感”。随着甲型H1N1流感蔓延多国,当前防控工作是重中之重,绝不可掉以轻心。     

这个春天,我们预防禽流感

流感病毒是流行性感冒病毒的简称,在分类上流感病毒属正粘病毒科,包括甲、乙、丙(也称A、B、C)3个型别。流感病毒可引发人类、禽类和动物类的“流行性感冒”,其中禽流感病毒是甲型流感病毒中的某些亚型(如H2N1、H9N2、H7N7等)。它们以鸡、鸭等禽类病毒贮存,宿主可以感染人类,早年已有若干先例。而此次出现的禽流感病毒主要为甲型流感病毒中的H5N1亚型。     

微科普

<正>科学松鼠会人的上呼吸道(鼻窦粘膜、咽喉、气管和支气管细胞)主要分布着α-2,6唾液酸受体。本来,人流感病毒(主要是H1,H2和H3亚型)对其亲和力较大,但突变后的禽流感病毒也具有了相当的亲和力,比如H5N 1和H7N 9型。这也是为什么现在H 7N 9禽流感让人类如此重视的原因之一。     

美疾控中心称美国的猪流感病毒为“杂交型”

世界卫生组织确认,美国和墨西哥出现了人感染猪流感（我国现称甲型H1N1流感）疫情,其中墨西哥为“重灾区”。美国疾控中心的消息报道说,经检测,这是A/H1N1型猪流感病毒的一个新型变异毒株;它包含禽流感、猪流感和人流感三种流感病毒的脱氧核糖核酸片断,同时拥有亚洲猪流感和非洲猪流感病毒的特征,因此是一种奇特的“杂交型”猪流感病毒。     

2009年甲型H1N1流感临床研究回顾

2009年全球爆发的甲型H1N1流感由一种源于猪流感病毒、致人急性呼吸道疾病的新型流感病毒所致.本文总结了甲型H1N1流感病毒感染病例的临床特征、治疗及研究展望.     

基于网络的流感病毒复制的研究

流感病毒是一种重要的呼吸道疾病病源,容易在人群中造成地方流行,给人类带来灾难和危害。利用A型流感病毒的4种毒株H3N2、H5N1、H7N7和H7N9的基因表达谱数据集,运用偏最小二乘法,分别构建了4个流感病毒感染复制的调控网络,挖掘了4个数据集下在病毒复制过程中发挥重要功能的基因,为进一步探索流感病毒在宿主细胞内的复制机制,揭示流感病毒的致病机理提供了可能。     

KMB17细胞培养流感病毒的实验研究

 探讨了流感病毒在KMB₁₇细胞上培养的可行性.将3株不同型别的流感病毒(A/Wisconsin/67/2005H3N2,A/New Caledonia/20/1999 H1N1和B/Malaysia/2506/2004)分别接种于KMB₁₇细胞上,在无血清的条件下于不同pH值和不同胰酶浓度的维持液中35℃培养72 h后收获病毒,测定血凝效价.实验结果表明,所用的3个毒株中的H3N2亚型毒株(A/Wisconsin/67/2005 H3N2)血凝效价明显高于另外的H1N1亚型(A/NewCaledonia/20/1999 H1N1)和B型(B/Malaysia/2506/2004)毒株.因此认为KMB₁₇细胞可作为培养流感病毒的备选培养基质.     

TritonX-100裂解流感病毒的研究

 在流感病毒高浓度条件下,寻找TritonX-100裂解流感病毒的适宜条件.在不同条件下用TritonX-100裂解WHO推荐用2007～2008年度流感病毒疫苗毒株制备的3个亚型的流感病毒,用电镜观察裂解程度,通过蔗糖密度梯度离心纯化抗原后免疫动物,以检测疫苗的免疫效果.结果表明当流感病毒的血凝效价是1:16382,裂解时间为24h时,TritonX-100在质量分数为1%可以完全地裂解B型,质量分数达到2%才可以完全裂解流感病毒H1N1和H3N2,这说明TritonX-100对A,B亚型毒株的裂解效果不同.由于每年都要更换毒株工人生产新的流感疫苗,因此应该对使用TritonX-100的裂解不同亚型的流感病毒条件进行研究,以保证裂解疫苗的质量.     

重症甲流感病毒肺炎影像学表现

探讨重症甲流感病毒肺炎的肺部CT、X线表现。收集甘肃省武威市凉州医院2009年9月至2009年11月收治的经病原学确诊的重症甲型H1N1流感病毒肺炎(甲流肺炎)患者14人,并结合文献资料报道的H7N9、H5N1等亚型重症相关报道资料,观察肺部CT及X线表现。重症甲流感病毒肺炎的肺部CT、X线表现主要为磨玻璃影、肺部实变影。重症甲流感病毒肺炎影像表现多灶、多态,转归迅速。     

Genome evolution of novel influenza A （H1N1 ） viruses in humans

The epidemic situation of A H1N1 flu arose in North America in April 2009, which rapidly expanded to three continents of Europe, Asia and Africa, with the risk ranking up to 5. Until May 13th, the flu virus of A H1N1 had spread into 33 countries and regions, with a laboratory confirmed case number of 5728, including 61 deaths. Based on IRV and EpiFluDB database, 425 parts of A H1N1 flu virus sequence were achieved, followed by sequenced comparison and evolution analysis. The results showed that the current predominant A H1N1 flu virus was a kind of triple reassortment A flu virus： （i） HA, NA, MP, NP and NS originated from swine influenza virus; PB2 and PA originated from bird influenza virus; PB1 originated from human influenza virus. （ii） The origin of swine influenza virus could be subdivided as follows： HA, NP and NS originated from classic swine influenza virus of H1N1 subtype; NA and MP originated from bird origin swine influenza virus of H1N1 subtype. （iii） A H1N1 flu virus experienced no significant mutation during the epidemic spread, accompanied with no reassortment of the virus genome. In the paper, the region of the representative strains for sequence analysis （A/California/04/2009 （H1N1） and A/Mexico/4486/2009 （H1N1）） included USA and Mexico and was relatively wide, which suggested that the analysis results were convincing.     

Orisin and future distribution of the new A （H 1N1 ） influenza virus emerging in North America in 2009

The origin of the new A （H1N1） influenza virus recently emerging in North America is a hot controversial topic of significance in disease control and risk assessment. Some experts claimed that it was an unusually mongrelized mix of human, avian and swine influenza viruses, while some others concluded that it was totally a simple re-assortment hybrid of two lineages of swine influenza viruses. Here the phylogenetic diversity of the viral PB1, PA and PB2 gene sequences using online web servers, and the results suggest that all the 8 genetic segments of the new virus were possibly from two lineages of swine influenza viruses, and one of the lineage was a mongrelized mix of human, avian and swine influenza viruses emerging in the world approximately 10 years ago. Considering the recent epidemiological trends of the new virus, we believe it will spread more widely in the world and persist long in human populations. It also could spread among swine populations. The future wide spreading of the new virus may coincide the disappearance of a subtype of previous human influenza A virus.     

Subtyping of type A influenza by sequencing the variable regions of HA gene specifically amplified with RT-PCR

The outbreak of a novel influenza A （H1N1） virus across the globe poses a threat to human health. It is of paramount importance to develop a rapid, reliable and inexpensive diagnostic procedure. Based on the bioinformatic information from public database, primers specific for influenza A virus surface protein haemagglutinin （HA） of several subtypes （including H1, H2, H3, H5, H7 and H9） were designed. Primer-specific PCR products were subiected to sequencing for accurately distinguishing H1 and H3 subtypes from others. This sequencing-based detection method will not only be applied to rapid detection and simultaneous subtype identification of new influenza A virus H1N1, but also provide the strategies to monitor other new types of influenza virus with explosive potential.     

姜黄素体外抗流感病毒H1N1、H3N2实验研究

目的：观察姜黄素提取物体外对流感病毒H1N1、H3N2的抑制作用。方法：用狗肾细胞（MDCK）观察姜黄素体外对A型流感病毒H1N1亚型、A型流感病毒H3N2亚型病毒的直接杀灭作用。结果：姜黄素最大无毒浓度为12．5g／L,对H1N1有效抑制浓度为6．25g／L,对H3N2有效抑制浓度为1．56g／L。结论：姜黄素提取物确有明显的抗H1N1、H3N2复制作用。     

Epidemiological and risk analysis of the H7N9 subtype influenza outbreak in China at its early stage

Dozens of human cases infected with H7N9 subtype avian influenza virus (AIV) have been confirmed in China since March, 2013. Distribution data of sexes, ages, professions and regions of the cases were analyzed in this report. The results showed that the elderly cases, especially the male elderly, were significantly more than expected, which is different from human cases of H5N1 avian influenza and human cases of the pandemic H1N1 influenza. The outbreak was rated as a Grade Ⅲ (severe) outbreak, and it would evolve into a Grade IV (very severe) outbreak soon, using a method reported previously. The H7N9 AIV will probably circulate in humans, birds and pigs for years. Moreover, with the driving force of natural selection, the virus will probably evolve into highly pathogenic AIV in birds, and into a deadly pandemic influenza virus in humans. Therefore, the H7N9 outbreak has been assumed severe, and it is likely to become very or extremely severe in the future, highlighting the emergent need of forceful scientific measures to eliminate any infected animal flocks. We also described two possible mild scenarios of the future evolution of the outbreak.     

H5N1亚型禽流感病毒血凝素基因的克隆与表达

 根据已知H5N1亚型禽流感病毒血凝素(HA)基因序列设计、合成克隆引物.自灭活的云南地方H5N1亚型病毒阳性临床组织样品中提取总RNA,反转录后采用高可信度DNA聚合酶(Pyobest^TMDNA Polymerase)扩增HA基因,采用Invitrogen定向表达系统(Champion^TMpET directional TOPO expression system)进行克隆表达,纯化获得N末端携带多聚组氨酸标签的重组HA,分子质量约78ku.采用阳性血清经免疫印迹及ELISA分析重组HA的免疫反应性,结果表明重组HA能与H5N1亚型病毒抗血清发生特异性结合,具有良好的免疫反应性.     

禽流感病毒H9亚型特异性抗原捕捉ELISA检测方法的研究

 采用禽流感病毒多克隆抗体及H9亚型特异性单克隆抗体,研究建立H9亚型特异性抗原捕捉ELISA检测方法,用于检测H9亚型禽流感病毒.优化了反应条件,确定了包被抗体、检测抗体及酶结合物的最佳工作浓度,对该方法的敏感性、特异性、重复性及稳定性分析,并与RT-PCR方法比较.通过使用该方法对野外样品进行检测.结果表明该方法敏感、特异,具有良好的重复性和稳定性,可用于检测临床样品、鸡胚培养物及细胞培养物中的H9亚型禽流感病毒.     

Experimental adaptation of an influenza H5 HA confers respiratory droplet transmission to a reassortant H5 HA/H1N1 virus in ferrets

Highly pathogenic avian H5N1 influenza A viruses occasionally infect humans, but currently do not transmit efficiently among humans. The viral haemagglutinin (HA) protein is a known host-range determinant as it mediates virus binding to host-specific cellular receptors. Here we assess the molecular changes in HA that would allow a virus possessing subtype H5 HA to be transmissible among mammals. We identified a reassortant H5 HA/H1N1 virus-comprising H5 HA (from an H5N1 virus) with four mutations and the remaining seven gene segments from a 2009 pandemic H1N1 virus-that was capable of droplet transmission in a ferret model. The transmissible H5 reassortant virus preferentially recognized human-type receptors, replicated efficiently in ferrets, caused lung lesions and weight loss, but was not highly pathogenic and did not cause mortality. These results indicate that H5 HA can convert to an HA that supports efficient viral transmission in mammals; however, we do not know whether the four mutations in the H5 HA identified here would render a wholly avian H5N1 virus transmissible. The genetic origin of the remaining seven viral gene segments may also critically contribute to transmissibility in mammals. Nevertheless, as H5N1 viruses continue to evolve and infect humans, receptor-binding variants of H5N1 viruses with pandemic potential, including avian-human reassortant viruses as tested here, may emerge. Our findings emphasize the need to prepare for potential pandemics caused by influenza viruses possessing H5 HA, and will help individuals conducting surveillance in regions with circulating H5N1 viruses to recognize key residues that predict the pandemic potential of isolates, which will inform the development, production and distribution of effective countermeasures.     

动物源性流感病毒与人流感流行

2009年4月.北美地区发生了人感染H1N1"猪流感"疫情,并且疫情呈现暴发漫延的态势.全世界关注的目光继1997年H5N1禽流感病毒感染人事件首次在中国香港发生,以及2003年以来包括15个国家和地区多起H5N1禽流感病毒感染人事件之后,再次聚焦到动物源性流感病毒上.是否动物流感病毒已经具备了引起人类疫情暴发的能力?是否新的一场大的人流感疫情就要暴发?为什么世界卫生组织又于4月30日为本次"猪流感"更名为"甲型H1N1流感"?对于流感暴发脚步的逼近我们是否做好了准备?本文对历史上人流感疫情暴发情况,动物流感病毒感染人事件的发生情况和流行病学特点,以及人流感疫情发生与动物流感病毒之间的关系等进行了总结与分析,以期更好地了解流感病毒在不同宿主之间的传播,为更加从容地面对这场突如其来的疫情,以及为疫情的预防和控制提供理论指导.为了能够准确反映这次流感的情况和国际对流感及流感病毒的统一命名法则,以及更加清晰并易于理解地表述清楚.我们建议将这次流感称作"北美流感"或"墨西哥流感".