人类流感病毒的多重RT-PCR方法分析

目的对人类流感病毒的多重RT-PCR方法的应用进行分析探讨。方法对40份疑似swineH1N1的临床标本采用多重RT-PCR方法进行检测,将检测结果进行统计分析,同时对该方法的特异性以及敏感性进行了评价。结果多重RT-PCR方法能够同时分析A型流感病毒M基因203bp、B型流感病毒M基因296bp。特异性实验表明多重RT-PCR法没有检测出存在非特异扩增的PCR产物,并且该方法在40份标本中检测出新甲型H1N1阳性标本4份（10.0%）,季节性H1N1亚型2份（5.0%）,H3N2亚型12份（30.0%）。结论多重RT-PCR方法具有快速、敏感性强、特异性强等优势,值得推广。     

甲型H1N1、季节性H1N1及H3N2亚型流感病毒的多重荧光RT-PCR方法的建立

目的构建一种可同时检测甲型H1N1、季节性H1N1、H3N2亚型流感病毒的多重荧光RT-PCR方法。方法根据不同流感亚型HA与NA的差异,设计五对引物及五条探针,构建合适的检测体系,分析特异性、灵敏度、可重复性、仪器适用性,并将其与常用检测试剂盒进行比对。结果本研究成功构建多重荧光RT-PCR检测体系,能同时检出甲型H1N1、季节性H1N1及H3N2亚型流感病毒,检测灵敏度为101~102拷贝/μl,可重复性好(变异系数为0.47%~4.03%),比常用检测试剂具有更好的荧光值水平,可适用于多种常见的荧光定量PCR仪。结论本方法所构建检测流感亚型体系具有较好的特异性、灵敏度、可重复性、仪器适用性,适用于各基层疾控及医疗机构。     

甲型H1N1流感确诊病例临床特点和治疗研究

目的分析甲型H1N1流感确诊病例的临床特征,探讨治疗方法,为防治提供依据。方法对229例甲型H1N1流感确诊病例的一般情况,临床表现,实验室检查,胸部X线,治疗方法进行统计、分析。结果甲型H1N1流感确诊病例平均年龄18.12岁,男：女为3.09：1,临床症状、体征中前三位分别是咳嗽157例、咽痛135例、咳痰50例和发热228例、咽部充血225例。白细胞总数降低40例、中性粒细胞和淋巴细胞降低占多数。血清酶升高：ALT22例、AST29例、LDH26例、CK45例、CKMB8例,血脂降低：CHO83例、TG108例。胸部X线异常6例,单侧肺炎1例,双侧肺炎5例。中药使用率100%,达菲使用率35.80%,抗生素使用率16.59%,其它抗病毒药物使用率13.10%。平均住院5.21d。结论甲型H1N1流感确诊病例以咳嗽、咽痛、咳痰、发热、咽部充血和肺部病变居多,CT检出肺部病变多,绝大部分血白细胞降低,心肌和肝脏最易受侵害,药物治疗效果好,病程较短。     

2009甲型H1N1流感病毒新进展

甲型猪源性流感病毒属正粘病毒科,其可引起猪、禽和人的上呼吸道感染。2009年3月在美国和墨西哥的流感样患者的呼吸道标本中检测出新的不能分型的甲型流感病毒,并经确认为甲型H1N1流感病毒（简称2009甲型H1N1流感病毒）。这种病毒在美国和墨西哥引起暴发流行,并蔓延到世界各地,引起一些感染者死亡。文中综述了2009甲型H1N1流感病毒在基因分型、抗原特征、传播特点、致病特点、治疗等方面与已知各流感病毒不同的特征。     

浅谈甲型H1N1流感

在人类繁衍发展的历史进程中,人与病原微生物的斗争总是周而复始,循环往复,但几乎每次都会以人类战胜病原微生物而告一段落,而后又会有新的病原微生物卷土重来,给人类造成一次又一次的灾难。2009年3月,一种新的流感病毒再次袭击人类,后经研究发现造成此次疫情的病毒株包含有猪流感、     

快速检测人新甲型H1N1、季节性和乙型流感病毒多重PCR方法的建立

目的 建立一种可以同时检测人类新甲型H1N1、季节性H1N1和H3N2亚型流感病毒及乙型流感病毒的多重PCR方法.方法 根据以上4种流感病毒的特异性基因设计4对引物,建立可同时扩增出4个流感病毒特异性片段的多重PCR的方法.用人的GAPDH基因作为内参判断标本来源和实施质量控制的依据.并进行敏感性、特异性和盲样检测评价实验.结果 该方法可同时扩增出流感病毒亚型的HA基因片段长度分别为106 bp(新甲型H1N1)、135 bp(季节性H1N1)、118 bp(季节性H3N2)、170 bp(乙型流感病毒).该实验检测新甲型H1N1(A/colifornia/07/ 2009)的敏感性为102copies/反应.特异性实验结果显示每对引物只检测对应的病毒,32份盲样检测结果特异性好.结论 该多重PCR方法可同时检测新甲型H1N1及季节性流感病毒,敏感性、特异性强,可用于人类流感病毒的鉴别分型检测.     

2009年新型甲型H1N1流感病毒血凝素基因进化分析

目的:探讨2009年新型甲（A）型H1N1流感病毒血凝素（HA）基因与世界各地不同年代分离的A/H1N1代表株HA基因的进化关系。方法:从NCBI数据库下载2009年新型A/H1N1亚型流感病毒的HA基因序列以及以往流行的人、猪和禽的A/H1N1亚型流感病毒参考序列,采用Molecular Evolutionary Genetics Analysis version 4.0（MEGA4.0）软件进行序列比对和构建系统进化树,并分别比较2009年新型甲型H1N1流感病毒HA基因与北美地区、欧洲地区、亚洲地区A/H1N1流感病毒HA基因编码蛋白的氨基酸序列。结果:不同时期人A/H1N1亚型流感病毒代表株HA基因进化分析显示：2009年新型A/H1N1流感病毒HA基因与1976~2007年北美地区分离的7株人A/H1N1亚型流感病毒具有较高的同源性,与欧洲和亚洲地区的同源性较低。不同种属间A/H1N1亚型流感病毒HA基因进化分析显示：2009年新型A/H1N1流感病毒HA基因与1998和2007年北美地区分离的A/H1N1猪流感病毒的HA基因进化关系较近,与欧洲及亚洲地区分离的A/H1N1猪流感病毒及A/H1N1禽流感病毒进化关系较远。氨基酸比对结果显示2009年新型A/H1N1流感病毒HA基因的重要抗原位点与北美地区分离的A/H1N1猪流感病毒相近,与欧洲和亚洲地区分离的A/H1N1猪流感病毒及人类流感病毒疫苗株相比变化较大。结论:2009年新型甲型H1N1流感病毒HA基因可能是北美地区甲型H1N1猪流感病毒长期进化并与该地区人A/H1N1流感病毒部分基因片段重排的结果,对人H1N1甲型流感病毒疫苗可能并不敏感。     

新型甲型H1N1流感病毒深圳分离株HA基因的克隆与进化

目的 分析2009年新型甲型H1N1流感病毒深圳分离株A/Shenzhen/71/09血凝素(Hemagglufinin,HA)基因序列,探讨其基因变异与分子进化.方法 采用RT-PCR技术扩增A/Shenzhen/71/09 HA基因片段并进行测序,利用Cluster和Mega3.1软件对不同亚型毒株HA基因核苷酸序列进行分析,绘制发育进化树. 结果 A/Shenzhen/71/09毒株HA基因包含HA1和HA2两个片段,长度分别约为1 032bp和669bp,HA蛋白裂解位点为VPSIQSR↓ GLF,不含连续碱性氨基酸.进化分析表明该病毒株与其它亚型病毒株遗传距离较远,而与深圳地区同期分离的其它H1N1毒株高度同源,同源性在99.0％以上.结论 新型甲型H1N1流感病毒HA基因不具备高致病流感病毒序列特征,深圳地区同期分离的毒株变异率低,但仍需密切关注新型甲型流感的传播情况.     

2009年新型甲型H1N1流感病毒聚合酶编码基因进化分析

目的:探讨2009年新型甲型流感病毒(A/H1N1)聚合酶PA、PB1和PB2编码基因的进化规律。方法:从NCBI流感病毒基因数据库下载2009年新型甲型H1N1流行株的PA、PB1和PB2聚合酶编码基因序列以及人、猪和禽流感病毒相应的参考序列,采用Molecular Evolutionary Genetics Analysis version 4.0（MEGA4.0）软件比对和修剪此次流行株的代表序列及所有参考株序列并构建系统树,再比对和修剪此次流行株的代表序列及人A/H1N1病毒各年代(1918~2008年)参考序列并构建系统树,同时比对此次流行株的代表序列及人A/H1N1各年代(1918~2008年)参考序列编码PB2蛋白的氨基酸序列。结果:不同地区分离的2009年新型甲型H1N1流感病毒的聚合酶PA、PB1和PB2编码基因均具有高度同源性,并聚集在一个独特的进化支上,与猪流感病毒对应基因接近。三者均与2005年美国爱荷华州分离的人A/H1N1病毒基因(A/Iowa/CEID23/2005/H1N1)具有高度的相似性。2009年新型甲型H1N1流感病毒、2005年美国爱荷华州流行的H1N1 (DQ889682)病毒PB2蛋白第627位氨基酸与禽类流感病毒相同,均为谷氨酸,而与其他人A/H1N1 (1918~2008年)病毒的赖氨酸不同。结论:2009年新型甲型H1N1流感病毒聚合酶基因可能来源于2005年美国爱荷华州分离的人A/H1N1病毒,禽流感病毒可能参与了聚合酶基因的重排过程。     

流感病毒快速检测法对甲型H1N1流感的临床诊断价值

目的探讨流感病毒快速检测法对甲型H1N1流感的临床诊断应用价值。方法对河南省平顶山市疾病预防控制中心2011年9月～12月之间收治的40例流感患者采取流感病毒快速检测法进行检测(A组),同时采取聚合酶链式反应进行检测流感病毒(B组),观察两组的检查率、敏感性与特异性。结果 A组流感病毒阳性率为67.5%,B组流感病毒阳性率为62.5%,两组病毒阳性检测率比较,差异无统计学意义(P>0.05);A组的假阳性表达率为15.0%(6/40),假阴性表达率为5.0%(2/40);而B组的假阳性表达率为10.0%(4/40),假阴性表达率为10.0%(4/40)。A组的敏感度为91.3%(21/23),特异性为64.7%(11/17);B组的敏感度为84.0%(21/25),特异性为73.3%(11/15),两组敏感度比较,差异有统计学意义(P<0.05)。结论采取流感病毒快速检测法在甲型H1N1流感的检测中具有较高的敏感性,能够满足临床中甲型H1N1流感的快速诊断,具有重要的临床诊断价值。     

Comorbid presentation of severe novel influenza A (H1N1) and Evans syndrome: a case report

One 22-month-old boy who was admitted for a fever lasting 6 days as well as a cough and wheezing lasting 2 days was reported. He was diagnosed with influenza A (H1N1, severe type), severe pneumonia, acute respiratory distress syndrome (ARDS), Evans syndrome and multiple organ failure. This is the first case of novel influenza A (H1N1) and Evans syndrome. The pathogenesis is still unknown.     

姜良铎教授治疗甲型H1N1流感病例分析(附3例报告)

目的总结姜良铎教授采用中药治疗甲型H1N1流感病例的有效经验。方法运用中医理论对甲型H1N1流感辨证论治,通过对不同病程阶段的症状、体征、方药归类,总结相应的证型特点和理法方药。结果 甲型H1N1流感可分为风热袭肺、痰热壅闭、毒伤肺络、津血外溢、气耗津伤等证候类型,以此辨证施治,归纳出有效的中医治疗方案。结论运用中医理论对甲型H1N1流感病程辨证分型,对症采取疏风清热、清肺化痰及祛湿清热、回阳救逆、益气养阴方法治疗,取得显著疗效。     

H7N9禽流感病毒实时荧光RT-PCR检测方法的建立与评价

目的应用Taq Man探针实时荧光RT-PCR技术,建立一种快速检测H7N9禽流感病毒的方法。方法针对H7N9禽流感病毒的HA和NA基因分别设计特异性引物和Taq Man-LNA探针,建立H7N9禽流感病毒特异性实时荧光RT-PCR快速检测方法。结果本研究建立的方法对检测H7N9禽流感病毒株的灵敏度达到10 PFU/ml。该方法特异性强,与其他亚型流感病毒株及呼吸道病原体无交叉反应。与对照方法相比,本研究方法的检测阳性符合率为99.07%,阴性符合率为100%,Kappa值为0.993。结论建立的检测试剂盒具有快速灵敏、结果准确可靠等优点,适用于H7N9禽流感病毒的分子生物学检测和监测。     

Comorbid presentation of severe novel influenza A (H1N1) and Evans syndrome: a case report

One 22-month-old boy who was admitted for a fever lasting 6 days as well as a cough and wheezing lasting 2 days was reported. He was diagnosed with influenza A (H1N1, severe type), severe pneumonia, acute respiratory distress syndrome (ARDS), Evans syndrome and multiple organ failure. This is the first case of novel influenza A (H1N1) and Evans syndrome. The pathogenesis is still unknown.

     

河南省18岁以下人群甲型H1N1流感抗体水平调查分析

目的 了解河南省18岁以下人群甲型H1N1流感抗体水平,为甲流防控提供相关依据。方法 采用微量血凝抑制试验对国家卫生部“甲型H1N1流感血清学横断面调查”的2020人(≤18岁)进行甲型H1N1流感抗体水平检测。结果 河南省18岁以下人群血清抗体阳性率为29.60%,13~18岁、7~12岁、0~6岁阳性率分别为45.70%、25.67%、23.71%,各年龄组阳性率比较差异有统计学意义(χ2=80.07,P<0.01)。城乡有差异(郑州市35.47%、其他市36.00%、农村23.67%)。接种疫苗人群抗体水平(63.89%)显著高于未接种人群(25.91%),χ2=117.18,P<0.01;有无流感样症状者的抗体阳性率差异无统计学意义(χ2=0.75, P>0.01)。结论 18岁以下人群具有一定的甲型H1N1流感抗体,但抗体水平较低,人群普遍易感,建议提高甲型H1N1流感疫苗接种率,进一步加强甲型H1N1流感防控措施。     

Hospitalized patients with novel influenza A (H1N1) virus infection: Shanghai, June–July 2009

Background From late May 2009, sporadic imported cases of novel influenza A （HIN1） were continuously confirmed in Shanghai, but there were few reports on its clinical presentation in China. The aim of the study was to investigate the demographic and clinical features of the laboratory-confirmed cases and the treatment with oseltamivir. Method We performed a retrospective study in the Shanghai Public Health Clinical Center （SHAPHC）, reviewing the medical records of the laboratory-confirmed patients derived from June 10 to July 20, 2009. Results A total of 156 cases were enrolled, of whom 152 had a history of recent travel. The mean age was 22.6 years and 89 cases （57.1%） were males. The most common symptoms were fever, cough, and sore throat, with children more likely to run a temperature above 38.5℃ than adults. The mean leucocyte count was 5.4×10^9/L, the mean neutrophil count 3.2×10^9/L and the mean lymphocyte count 1.4×10^9/L. Other findings included a normal range or elevated level of C-reactive protein （CRP） and glutamic-pyruvic transaminase and a normal or decreased level of prealbumin; the levels of prealbumin and CRP were significantly lower in the children than in the adults. Fifty-two patients had abnormal chest CT results, with small unilateral or bilateral pulmonary infiltrates, axillary and mediastinal lymphadenopathy and local pleural thickening, while no cases showed symptoms of hypoxia. All the patients received oseltamivir and recovered without complications, but the duration of fever and virus shedding were significantly longer in the children than in the adults. Conclusions Travel-related circulation may be an important reason for the H1N1 epidemic in the non-epidemic areas, and the virus caused mild respiratory symptoms. The infection in children was more severe in terms of prealbumin levels, temperature, the duration of fever and virus shedding. Oseltamivir was effective for H1N1, but more effective in the adults than in the children.     

温州市2009-2010年甲型H1N1流感疫情与流感监测分析

目的分析温州市2009-2010年甲型H1N1流感的流行病学特征和流感监测结果,为科学防控甲型H1N1流感和流感大流行提供依据。方法收集中国疾病预防控制信息系统中2009-2010年温州市甲型H1N1流感报告信息和流感监测信息并进行描述流行病学分析。结果 2009-2010年温州市共报告甲型H1N1流感4 288例,其中2009年报告3 959例,报告发病率为51.28/10万,死亡4例,病死率为0.10%;2010年报告329例,报告发病率为4.22/10万,死亡2例,病死率为0.61%。首发病例为从美国回国的留学生。病例主要集中在0～29岁,占全部病例数的89.51%;流感样病例占门诊就诊总数百分比(ILI%)从2009年第30周开始超过2007和2008年水平,甲型H1N1流感疫情在2009年9～11月达到流行高峰。经济相对发达的地区发病率高于其他地区。结论儿童、青少年和学生群体是甲型H1N1流感防控重点人群,应加强流感哨点监测,重点防控甲型H1N1流感聚集性病例和重症危重病例。     

Sixty-two severe and critical patients with 2009 influenza A (H1N1) in Shanghai, China

Background Pandemic influenza A (H1N1) emerged rapidly in China in May 2009. Preliminary comparisons with seasonal influenza suggest that pandemic 2009 influenza A (H1N1) disproportionately affects younger ages and causes generally mild disease. To characterize disease progress, comorbidities, and treatment outcomes among consecutive severe and critically ill patients in a hospital served as a reference center for the care of patients with H1N1 in Shanghai,China.Methods A retrospective study on 62 severe and critically ill patients with 2009 influenza A (H1N1) was conducted in Shanghai Public Health Clinical Center. Demographic data, symptoms, comorbidities, disease progression, treatments,and clinical outcomes were collected for analysis.Results Sixty-two severe or critically ill patients were admitted to the hospital with confirmed 2009 influenza A (H1N1) infection. The median age of the study cohort was 40 years old with a range from 18 years to 75 years, and 67.7% were males. All patients presented with fever and respiratory symptoms. At presentation, 34 patients (54.8%) had comorbidities such as smoking (29.0%), hypertension (29.0%) and hepatitis B virus infection (9.7%). The median time from symptom onset to hospital admission was 6 days (interquartile-range 3-14 days) and 23 critically ill patients were admitted to Intensive Care Unit after admission. All the patients received neuraminidase inhibitors (oseltaminir), while 60 patients (96.7%) were treated with antibiotics, and 39 (62.9%) with corticosteroids. Twenty-three critical cases received noninvasive mechanical ventilation on the first day of admission, and 3 of them ultimately required invasive ventilation.Four death reports (6.5%) were filed within the first 14 days from the onset of critical illness with the primary causes of severe acute respiratory distress syndrome, hypoxemia, or complications, secondary infection and sepsis,pyopneumothorax and stroke.Conclusions Severe illness from 2009 influenza A (H1N1) infection in Shanghai occurred among young individuals. Critical cases were associated with severe hypoxemia, multisystem organ failure, and a requirement for mechanical ventilation. Most patients had a good prognosis.     

Anti-pandemic influenza A (H1N1) virus potential of catechin and gallic acid

Background

The pandemic influenza A (H1N1) virus has spread worldwide and infected a large proportion of the human population. Discovery of new and effective drugs for the treatment of influenza is a crucial issue for the global medical community. According to our previous study, TSL-1, a fraction of the aqueous extract from the tender leaf of Toonasinensis, has demonstrated antiviral activities against pandemic influenza A (H1N1) through the down-regulation of adhesion molecules and chemokine to prevent viral attachment.

A型人流感H5N1病毒进化关系分析

目的:探讨A型人流感与禽流感H5N1病毒的基因组进化关系.方法:拼接99株A型流感H5N1病毒全基因组编码区序列,进行比对分析,构建基因组进化树.结果:A型人流感H5N1病毒分布在3个进化枝,与枝内禽流感病毒具有高度同源性.结论:虽然不同进化枝的A型人流感病毒来源均为禽源性的,但是不同进化枝之间的同源性可存在较大差异.