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目的 了解2009年度甲型H1N1流行性感冒(流感)病毒的检测情况和血凝素(HA)基因变异情况.方法 选择国家级流感监测哨点医院以及暴发疫情的疫点,采集流感样病例的鼻咽拭子标本,通过实时(RT)-PCR进行病毒分型及甲型H1N1流感病毒检测,对阳性标本采用狗肾细胞(MDCK)进行病原分离,采用红细胞凝集试验测定病毒效价,用血凝抑制实验进行型别鉴定,通过RT-PCR扩增毒株HA1片段的基因并进行序列测定,利用生物信息学技术进行序列分析.结果 共检测咽拭子样本996份,其中核酸检测阳性病例包括甲型H1N1 337份,季节性H1N1亚型1份,季节性H3N2亚型67份,B型12份,流感核酸检测阳性率为41.87%,其中甲型流感核酸检测阳性率为33.84%.分离出甲型H1N1病毒36株,选择18株.测序成功的10株甲型H1N1流感病毒在多个氨基酸位点发生变异,与疫苗株A/California/07/2009(H1N1)比较,有6个位点发生突变,其中1个位点位于抗原决定簇的B区.结论 2009年度分离到的流感病毒株中以甲型H1N1为绝对的优势毒株,毒株的血凝素基因与世界卫生组织(WHO)提供的疫苗株相比有变异,与疫苗株相比,抗原决定簇B区有改变,但关键位点第222位没有变化. 相似文献
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目的 研制检测靶位针对甲型流行性感冒(流感)病毒核蛋白的甲型流感病毒抗原检测试剂盒,建立能检测甲型流感病毒所有亚型的方法.方法 采用双抗体夹心法检测标本中的甲型流感病毒,建立甲型流感病毒抗原ELISA方法,并对该试剂盒的灵敏性、特异性、精确性、稳定性进行测试和临床考核.结果 甲型流感病毒抗原ELISA试剂盒核蛋白检测下限为7.63 ng/Ml,灵敏度是血球凝集方法的256倍,是美国Quickdel公司胶体金产品的16倍;与乙型流感病毒、呼吸道合胞病毒、呼吸道腺病毒、副流感病毒Ⅰ/Ⅲ型、肺炎支原体、鸡新城疫病毒、鸡法氏囊病毒、鸡传染性支气管炎病毒均无交叉,特异度为100%;批内、批间变异系数(CV)值均<15%,符合国家标准;在4℃以下的稳定性可达1年,能通过37℃7 d加速试验.可检测亚型分别为H1N1、H3N2、H5N1和H9N2的甲型流感病毒.人流感病毒临床试验表明,与常规细胞培养方法比较,阳性符合率为93.44%,阴性符合率为99.31%;禽流感病毒临床试验表明,与常规细胞培养方法比较,阳性符合率为95.45%,阴性符合率为98.09%.结论 甲型流感病毒抗原ELISA试剂盒灵敏性、特异性强,可用于人甲型流感病毒和禽流感病毒感染的流行病学调查. 相似文献
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目的 检测和分析2009年广东省甲型流行性感冒(流感)暴发流行期间首株甲型H1N1流感病毒血凝素(HA)基因.方法 对2009年广东省首例确诊的甲型H1N1流感患者的咽拭子进行病毒分离,取细胞培养上清液提取病毒核酸,采用HA基因的特异性引物进行RT-PCR,PCR产物进行克隆、测序和同源性分析.结果 获得2009年广东省首株甲型H1N1流感病毒的HA基因,大小为1710 bp,命名为A/GuangzhouSB/01/2009(H1N1)HA,GenBank登录号为GQ268003.与疫情发源地近期报告的277株甲型H1N1流感病毒的HA基因比对,同源性为99.0%~99.8%;其中与美国报告的病毒株的同源性高达99.8%,与患者发病前曾到美国旅游的流行病学史一致.与25株中国季节性甲型H1N1流感病毒的HA基因比对,同源性为72.3%~85.6%.结论 2009年广东省甲型流感暴发流行期间首株甲型H1N1流感病毒与目前流行的甲型H1N1流感病毒同源性高,与中国季节性甲型H1N1流感病毒同源性较低. 相似文献
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甲型(H1N1)(2009)流感病毒三重RT-PCR快速检测方法的建立 总被引:1,自引:0,他引:1
目的建立一种早期快速检测甲型(H1N1)(2009)流感流行病毒株的方法。方法将2009年分离的甲型(H1N1)流感流行病毒株基因与以前的流感病毒株进行序列比对,找出其特异的基因序列,设计针对甲型(H1N1)(2009)流行株的血凝素(HA)、神经氨酸酶(NA)、核蛋白(NP)基因的三对特异引物,采用RT-PCR同时扩增三条目的片段,通过琼脂糖凝胶电泳进行检测。结果此方法对临床标本的阳性检出率为71%。结论采用三重PCR同时扩增甲型(H1N1)(2009)流感流行病毒的三段特异序列既缩短检测时间又提高了检测特异性,无交叉反应,是一种有效可行的快速检测甲型(H1N1)(2009)流感流行病毒株的方法。 相似文献
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目的分离甲型HIN1流感病毒,分析福建省首例病毒分离株全基因组序列和遗传特征,为研究病毒进化、致病性、流行规律提供科学依据。方法采用MDCK细胞和Real—time PCR法进行病毒分离、鉴定;提取病毒RNA,通过RT-PCR扩增其8个基因片段,测定核苷酸序列,利用生物信息软件拼接全基因组序列;分析重要基因位点,利用GENBANK中相关序列对首例病毒分离株A/Fujian/01/2009(H1N1)进行基因进化树分析。结果从82例甲型H1N1流感确诊病例标本中分离出50株甲型H1N1流感病毒,第一代分离阳性率60.98%。在福建省首次获得甲型H1N1流感病毒株及全基因组序列。基因组序列分析证明:该毒株与2009年大流行株高度同源,其基因组存在四源重组现象;氨基酸位点分析其对达菲药物敏感,对金刚烷胺类药物耐药;相对于猪流感代表株A/Swine/Iowa/15/1930(HIN1)存在6个HA抗原决定簇位点变异。结论MD—CK细胞对甲型H1N1流感病毒具有较高敏感性;福建省首例甲型H1N1流感病例分离病毒株与北美流行株高度同源;相对于以往古典型猪流感代表株出现了HA蛋白抗原性漂移;为今后进一步开展甲型H1N1流感病毒分子生物学研究奠定基础。 相似文献
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目的 明确一株人甲型H1N1流行性感冒(流感)病毒的遗传背景,建立流感病毒反向遗传的平台.方法 对分离自汕头市儿童的一株人甲型H1N1流感病毒进行空斑纯化;利用甲型流感病毒通用引物,对该病毒全基因组的8条片段(PB2、PB1、PA、HA、NP、NA、M和NS)进行RT-PCR全长克隆、DNA测序及初步生物信息学分析;将其8条全长基因组片段分别插入反向遗传通用载体,构建人甲型H1N1流感病毒的反向遗传系统.结果 RT-PCR克隆得到该H1N1毒株的8条全长片段,经测序分析确认该毒株的基因序列,该毒株与2006年至2008年分离的人甲型H1N1流感病毒具有很高的同源性,未出现奥司他韦或金刚乙胺抗药性的遗传突变.PCR与测序证实,插入该菌株8个全长基因组片段的载体序列完全正确.结论 成功构建了该毒株的感染性克隆.获得了该毒株全基因组序列,为明确病毒遗传信息、提供流行病学监测数据、建立流感病毒反向遗传平台奠定了研究基础. 相似文献
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目的 探讨2009年广东省新型甲型H1N1流行性感冒(流感)病毒神经氨酸酶(NA)基因的进化及NA基因编码蛋白抗原性、酶活性位点、糖基化位点变异情况.方法 从2009年广东省新型甲型H1N1流感患者中分离到病毒毒株共69株,提取病毒总RNA,RT-PCR扩增NA基因,并测序分析;同时从美国国立生物技术信息中心基因库检索获得 52株不同年代、不同地域甲型流感病毒NA基因序列,用MEGA 4.0软件进行基因进化分析和氨基酸序列分析.结果 2009年广东省新型甲型H1N1流感病毒NA基因与禽H5N1流感病毒同源性较高,为87.1%,潜在抗原位点氨基酸分布相同;所有毒株的酶活性中心位点高度保守;具有8个糖基化位点,其中5个位点有不同程度的氨基酸替换,但与2001年禽H5N1毒株的糖基化位点的氨基酸相同.结论 2009年广东省新型甲型H1N1流感病毒NA基因与禽H5N1流感病毒高度同源,与NA抑制剂的特异性结合位点未发生变化. 相似文献
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目的分离目前流行的甲型H1N1流感病毒,对其进行基因序列测定和遗传变异分析,为研究病毒进化、致病性、流行规律及防治提供科学依据。方法采用MDCK细胞、SPF鸡胚和RT-PCR方法对甲型H1N1流感病毒进行分离、鉴定,并进行核苷酸序列测定和分子进化分析。结果从疑似甲型H1N1流感临床标本中分离得到1株甲型H1N1流感病毒,命名为长春株(A/Changchun/01/2009(H1N1)),基因序列及分子进化分析显示该毒株与2009年大流行的甲型H1N1毒株高度同源,属于流行分支2。相对流行分支1(北美流行株A/California/07/2009(H1N1)),PB2、HA、NA分别出现2个、7个和4个氨基酸的差异。结论甲型H1N1流感病毒长春株与2009年大流行的甲型H1N1毒株高度同源,属于流行分支2,与流行分支1相比存在变异。 相似文献
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目的:建立一种检测甲型H1N1、季节性H1N1、H3N2与乙型流感病毒的SYBR GreenⅠ实时荧光PCR方法。方法:设计针对甲型H1N1、季节性H1N1、H3N2与乙型流感病毒HA基因的引物,分别进行SYBR GreenⅠ实时荧光定量PCR反应,同时进行熔解曲线和Tm值分析,并作灵敏度和重复性试验。 结果:结果表明该方法与H5、H7、H9亚型流感病毒及副流感病毒1、2、3型均无交叉反应。构建的荧光定量标准曲线循环阈值与模板浓度呈良好的线性关系,扩增效率为:95.588%,检测灵敏度为101copies/反应体系,结果重现性好。成功地验证性检验了32份盲样标本。结论:本研究建立的SYBR GreenⅠ实时荧光PCR方法敏感、不需荧光标记探针、成本低及高通量,能用于人类流感病毒的分型检测。 相似文献
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两种新发现的脑炎病毒 总被引:1,自引:0,他引:1
1994年爆发了两次能感染马和人类的一种新的病毒性疾病 ,引起昆士兰和澳大利亚 2 3匹马和 3人感染 ,其中一人死于爆发的呼吸道疾病 ,另一人死于脑炎 ,第三位患者出现流感样症状并完全康复 ,引起该病的病毒最初称为马麻疹病毒 (equinemorbillivirus ,EMV) ,后命名为Hendra病毒。 1998年 9月下旬至1999年 6月中旬在马来西亚爆发的一种病毒性脑炎 ,发病例数高达 2 65例 ,其中 10 5例死亡。绝大多数患者是养猪场或屠宰场工人。 1999年 3月 10日~ 19日在新加坡 1个屠宰场的 11名工人出现脑炎或不典型肺炎的临床表现… 相似文献
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Dengue is a debilitating disease that poses a perpetual threat to human health and increases the global economic burden every year. Despite advances in medical sciences, dengue virus(DENV) infects approximately 200 million people every year. To date, no effective antiviral is valiable to treat DENV in individuals despite great efforts in accomplishing these goals. Numerous approaches have been used in the search for dengue antiviral like screening of combinatorial compounds against DENV enzymes and structurebased computational discovery. In recent years, investigators have turned their focus into medicinal plants, trying to identify compounds that can be used as dengue antiviral. Nature represents a great reservoir of potential substances that can be explored with the aim of discovering new drugs that can be either used directly as pharmaceuticals or can provide drug leads, which can be scrutinized further for the development of new anti-dengue natural product. Many previous investigations have dealt with numerous plant extracts or bioactive principles for their antiviral property as they normally considered being safer when compared to synthetic drugs. Andrographis paniculata belongs to family Acanthaceae and is generally known as ‘king of bitters'. Diverse bioactive compounds from this plant such as diterpenes, flavonoids, xanthones, noriridoides and other miscellaneous compounds have exhibited their potential as therapeutics for various chronic as well as infectious diseases. This review is based on literature review on scientific journals, books and electronic sources, which highlights the pathogenesis of DENV and describe an assortment of bioactive principles that have been possessing antiviral potential, which include dengue and discuss the therapeutic efficacy and mechanism of action of Andrographis paniculata. However, a detailed and more comprehensive clinical trial on mammalian tissues and organs is needed in future studies. 相似文献
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Calderwood MA Venkatesan K Xing L Chase MR Vazquez A Holthaus AM Ewence AE Li N Hirozane-Kishikawa T Hill DE Vidal M Kieff E Johannsen E 《Proceedings of the National Academy of Sciences of the United States of America》2007,104(18):7606-7611
A comprehensive mapping of interactions among Epstein-Barr virus (EBV) proteins and interactions of EBV proteins with human proteins should provide specific hypotheses and a broad perspective on EBV strategies for replication and persistence. Interactions of EBV proteins with each other and with human proteins were assessed by using a stringent high-throughput yeast two-hybrid system. Overall, 43 interactions between EBV proteins and 173 interactions between EBV and human proteins were identified. EBV-EBV and EBV-human protein interaction, or "interactome" maps provided a framework for hypotheses of protein function. For example, LF2, an EBV protein of unknown function interacted with the EBV immediate early R transactivator (Rta) and was found to inhibit Rta transactivation. From a broader perspective, EBV genes can be divided into two evolutionary classes, "core" genes, which are conserved across all herpesviruses and subfamily specific, or "noncore" genes. Our EBV-EBV interactome map is enriched for interactions among proteins in the same evolutionary class. Furthermore, human proteins targeted by EBV proteins were enriched for highly connected or "hub" proteins and for proteins with relatively short paths to all other proteins in the human interactome network. Targeting of hubs might be an efficient mechanism for EBV reorganization of cellular processes. 相似文献
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Residual risk of transfusion transmitted human immunodeficiency virus, hepatitis B virus, hepatitis C virus and human T lymphotrophic virus 总被引:4,自引:0,他引:4
BACKGROUND: The risk of transfusion transmitted viral infection is now so low that mathematical modelling is required to estimate the residual risk. The first national viral risk estimates for hepatitis B virus (HBV), human immunodeficiency virus (HIV) and hepatitis C virus (HCV) were recently published by the Australian Red Cross Blood Service. Using several refinements to the original methodology, as well as an additional 2 years of data, new risk estimates have been derived. METHODS: Viral screening data for Australian donors for 2000/2003 were retrospectively analysed. The data were applied to three published models to estimate the residual risk of transmitting HIV, HBV, HCV or human T lymphotrophic virus (HTLV) by blood transfusion in Australia. RESULTS: Applying the three models to HBV, HIV and HCV, three point estimates of the residual risk per unit were calculated for each virus. The median point estimates were 1 in 1,339,000 for HBV, 1 in 1 in 7,299,000 for HIV, and 1 in 3,636,000 for HCV. Although the HTLV risk could not be equivalently calculated because of the lack of incident infection it was estimated to be considerably less than 1 in 1,000,000 using a separate method. CONCLUSIONS: The most current and accurate estimate of residual risk of viral transmission in Australia has been provided in the present study. The residual risk in Australia is exceptionally small, continuing to decrease and is generally less than European or US risk estimates. These new estimates demonstrate that for viral transmission the Australian blood supply is amongst the safest in the world, and provide a basis for evaluating the cost benefit of future viral testing methodologies. 相似文献
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谭友文 《世界华人消化杂志》2011,(15):1614-1619
从全球范围看,乙型肝炎病毒(hepatitis B virus,HBV)和丙型肝炎病毒(hepatitis C virus,HCV)重叠感染估计约有700-2000万人口感染.重叠感染和单一HBV或HCV感染比较,更易发展为肝硬化、肝细胞癌甚至肝衰竭的比例也高,HBV和HCV重叠感染可有四种不同的临床模式,即HCV活动... 相似文献