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1.
目的 对广州市流感监测人群呼吸道标本进行甲型H1N1流感病毒分离和测序,并对耐药基因突变和遗传进化特征进行分析。方法 2017年1月-2018年3月,对4家流感监测医院的5831份监测标本进行甲型H1N1流感病毒核酸检测,将阳性标本接种MDCK细胞进行流感病毒分离和测序,应用DNA Star7.1软件和MEGA 4.0软件对耐药基因进行测序分析。结果 5831份监测标本中检出甲型H1N1核酸阳性标本222份,检出率为3.81%,其中分离甲型H1N1流感病毒61株,分离率为1.05%。有2株病毒分离株出现了H274Y突变,突变率为3.28%。所有病毒分离株均发生了S31N突变,突变率为100%,同时另有2株出现V27A突变,突变率为3.28%。部分2017年毒株和2018年毒株已形成于A/Michigan/45/2015疫苗株分支之外的另一独立小分支。结论 2017年广州市流感监测病例中监测到2株H1N1流感病毒分离株为奥司他韦耐药突变株,且该耐药株均位于当前疫苗株流行分支以外的独立分支,有必要对广州市甲型H1N1流感病毒进行持续监测。  相似文献   

2.
From March through June 1977 a total of 31 influenza A (H3N2) viruses were isolated from students with respiratory disease who were seen at the student health service on the Berkeley campus of the University of California, and 32 influenza A (H3N2) viruses were isolated from persons who participated in a city-wide febrile respiratory disease surveillance program in Seattle. The antigenic specificity of the hemagglutinin was determined for each isolate by hemagglutination inhibition testing with sera from ferrets infected with prototype strains A/Victoria/3/75 and A/Texas/1/77. In each of the three months, April, May and June, A/Victoria/3/75-like and A/Texas/1/77-like viruses were identified among isolates from both communities, and the numbers of isolates of the two antigenic variants from patients seen with influenza-like illnesses were similar. The findings emphasize the need to examine multiple isolates even from within single communities to determine the antigenic specificity of current strains of influenza virus.  相似文献   

3.
During 1981, the A/Brazil/11/78-like strains of influenza virus that had been prevalent from 1978 to 1980 were displaced by a new set of heterogeneous, but closely related, variants (reference strain, A/England/333/80). Genomic analysis revealed that these new variants were almost exclusively nonrecombinant H1N1 viruses, i.e., they contained no genes of H3N2 origin. However, a few recombinant viruses containing the new variant HA and genes of H3N2 origin were identified. Antigenic analysis of H3N2 viruses indicated that they were also heterogeneous. The majority of these virus isolates were antigenically intermediate between A/Texas/1/77 and A/Bangkok/1/79, but additional variants were detected. Genomic analysis revealed that the H3N2 viruses isolated in the winter of 1980-81 were quite similar to H3N2 viruses isolated from 1977-79 in their T1 oligonucleotide maps. No H1N1 genes were detected in H3N2 virus isolates. Comparison of pairs of oligonucleotide maps of total virus RNA indicated that a similar rate of genetic change had occurred for nonrecombinant H1N1 viruses, for recombinant H1N1 viruses, and for H3N2 viruses and that, in general, pairs of viruses exhibited increasing numbers of changes in their oligonucleotide maps as the time interval between isolation of the viruses increased.  相似文献   

4.
目的 从分子流行病学角度对盐城市2015-2017年流行的甲型H3N2流感病毒血凝素(hemagglutinin,HA)和神经氨酸酶(neuraminidase,NA)基因的分子特征进行研究。方法 收集盐城市2015-2017年流感监测哨点医院以及流感爆发点采集的3 891例流感样病例标本进行核酸检测和病毒分离,从中筛选出20株经血凝抑制试验(hemagglutination inhibition,HI)验证为甲型H3N2亚型的毒株,采用反转录聚合酶链式反应方法扩增其HA1和NA基因并进行测序。结果 2015-2017年流行的20株甲型H3N2毒株HA1和NA基因各自分支的聚类关系基本一致。盐城市A/Jiangsu-YC/1474/2015、A/Jiangsu-YC/1725/2015两株分离株与疫苗株A/Switzerland/9715293/2013和A/Hongkong/4801/2014的进化距离较近,而其余18株分离株与国内部分省市相近年份毒株亲缘关系相近,并与疫苗株A/Switzerland/9715293/2013和A/Hongkong/4801/2014进化距离较远。该20株毒株HA1基因编码区抗原表位、受体结合位点和糖基化位点均发生了一定程度变异。从基因变异角度进行分析,疫苗株A/Switzerland/9715293/2013对盐城地区流感流行的保护效果要弱于疫苗株A/Hongkong/4801/2014。结论 2015-2017年盐城市流行的甲型H3N2毒株HA1和NA基因已逐渐发生变异,可能导致流感病毒发生实质性的抗原性漂移,降低与流感疫苗株的匹配度,减弱流感疫苗的保护作用。  相似文献   

5.
目的 了解2017年云南省流感病毒的流行情况和甲型H1N1流感病毒的血凝素(HA)基因分子特征。方法 对2017年云南省流感监测哨点医院采集的21 672份标本使用real - time RT - PCR方法检测流感病毒,核酸阳性标本使用MDCK细胞进行病毒分离,采用血凝素凝集试验及抑制试验进行病毒抗原性分析。选取12株抗原性变异较大的甲型H1N1流感毒株进行基因测序,使用MEGA软件分析其HA基因特征。结果 2017年共检出流感核酸阳性标本2 372份,分离出流感毒株1 180株,全年共有2个流行高峰,H3亚型和甲型H1N1亚型交替成为优势株,BY亚型和BV亚型共同流行。毒株分离高峰与ILI%高峰基本一致。甲型H1N1流感病毒HA基因无明显变异,疫苗株仍具有保护作用。结论 继续加强监测力度,提高监测敏感性并做到及时预警,控制流感流行风险。  相似文献   

6.
天津市儿童流行性感冒病原学检测分析   总被引:7,自引:0,他引:7       下载免费PDF全文
目的:对天津市2001-2002年流行性感冒(流感)患儿进行病原学分析。方法:用狗肾传代(MDCK)细胞和鸡胚双腔法进行流感病毒分离。用鸡红细胞和人O型红细胞凝集试验证实病毒的存在,用红细胞凝集抑制试验进行型和亚型的鉴定。结果:2001年10月至2002年3月共采集14岁以下流感样患儿咽拭子标本238份,发离出64株流感病毒,检出率为26.9%;其中A(H3N2)亚型42株,占检出数的65.6%;A(H1N1)亚型13株,占20.3%;B型9株,占14.1%,所分离的64株流感病毒都具有适应于MDCK细胞株生长及与人O型红细胞凝集良好的生物学特性,转种鸡胚后,绝大多数(62/64)能适应鸡胚生长;但64份阳性咽拭子标本液直接接种鸡胚,仅有3份血凝阳性,此外,96.4%(53/55株)A型流感病毒均可由O相转为D相,有2株A(H3N2)为O相特征,B型流感病毒均为O相特征,结论:天津地区流感病毒存在A(H3N2)、A(H1A1)和B三个型,以A(H3N2)为优势流行型。  相似文献   

7.
目的 了解1998-2004年北京地区婴幼儿中流行的A3(H3N2)亚型流感病毒血凝素基因HA1区的基因变异特点。方法 提取该期间采集的呼吸道标本中H3N2亚型流感病毒的RNA,经逆转录多聚酶链反应扩增得到HA1区基因片段。通过目的基因的克隆、测序或PCR产物直接测序,进行序列分析。结果 19982004年问北京地区婴幼儿中流行的H3N2亚型流感病毒血凝素HA1区基因均为987bp,编码一个含329个氨基酸的蛋白质。这些H3N2亚型流感病毒血凝素HA1区的核苷酸和氨基酸同源性分别在95.5%~100.0%和93.0%~100.0%之间,分离年份越近同源性越高。在HA1区有7个糖基化位点非常保守,分别位于8、22、38、63、126、165和285位氨基酸。与1997年以前的毒株相比,1997年以后的毒株均在122和133位增加了一个糖基化位点。自1999年以后的毒株均在144位增加了一个糖基化位点。每年流行的毒株都较前一年的毒株出现了位于不同抗原决定簇或者受体结合位点的氨基酸替换。进化分析显示,每年分离的毒株均与以前的毒株出现了不同程度的变化,不断出现新的进化分支。结论 1998—2004年间北京地区婴幼儿中流行的H3N2亚型流感病毒持续不断地发生着点突变,导致抗原性不断发生漂移。加强监测和密切关注其变异动向对防控流感流行有极其重要的意义。  相似文献   

8.
In April-May 1980, two independent outbreaks of influenza-like illness occurred in Leningrad among children''s-home children aged from 3 months to 2 years (of 68 children under observation, 50 became ill) and among boarding-school pupils aged 15-17 years (of 50 pupils under observation, 13 became ill). A total of five influenza A virus strains were derived from one clinically healthy and three affected children of the children''s home. Similar viruses were obtained from one affected boarding-school pupil and from an infected woman aged 24 years (a sporadic case within a household). On the basis of laboratory findings, all these seven strains were identified as influenza A H2N2 subtype strains. Six of the affected children showed significant seroconversion only to H2 haemagglutinin from February to May 1980. Type A influenza H2N2 virus was isolated from three persons, including the sporadic case, who also showed significant seroconversion to H2 haemagglutinin. H2N2 influenza A virus was isolated on two occasions, at a 7-day interval, from the girl N. Ju. Laboratory findings obtained from the study of the viruses isolated using up-to-date immunological and molecular-biochemical techniques enable us to conclude the following. The A/Leningrad/80 isolates belong to H2N2 sero-subtype. The viruses isolated are similar but not identical to the A/Singapore/I/57 reference strain in details of polypeptide and gene composition.  相似文献   

9.
目的 对1998年浙江省的H3N2流感流行进行溯源研究.方法 采用RT-PCR扩增浙江省1998年3株H3N2流感流行代表株的全基因组序列,并与GenBank上1995-1998年世界其他地区H3N2流感流行株进行比较;同时,采用交叉血凝抑制实验,计算各毒株间的抗原比.结果 HA基因进化树表明,1998年浙江省H3N2优势流行株A/Zhejiang/11/98、A/Zhejiang/18/98与1995-1996年世界各地以及1997年我国大陆的H3N2流行株间存在显著差异,在进化树上虽与A/Sydney/5/97同属一簇,但和美国纽约以及中国香港1997年后期流行株更为接近.在HA1、NA和MP基因上,A/Zhejiang/18/98与香港1997年后期流行株同源性最高,而在PA、HA和NS基因上,与纽约流行株的遗传距离也小于A/Sydney/5/97.A/Zhejiang/18/98与香港或纽约株在HA1区仅存在1~3个位点的氨基酸残基不同,而与A/Sydney/5/97存在7个位点的氨基酸残基差异,其中3个位点于抗原决定簇区.各毒株间的交叉血凝抑制实验表明A/Zhejiang/18/98与A/Sydney/5/97的抗原比已达2.0,提示二者在抗原性上存在一定差异.此外,1997-1998年H3N2各地流感流行的起始时间序列,也显示了该次流感传播的可能途径.结论 浙江省1998年H3N2流感的流行很可能是由1997年底H3N2新型流感变异株经纽约和香港输入中国大陆所导致.  相似文献   

10.
In order to conduct a survey of influenza viruses entering Japan via travellers arriving by airplanes, gargle solutions were collected from passengers who reported to the quarantine station of Nagoya International Airport complaining of respiratory symptoms. From 504 samples collected between August 1996 and March 1999, 30 influenza virus strains were isolated. Twenty-eight of the isolates were influenza A (H3N2) viruses and two were influenza B viruses. No H1N1 virus was isolated. Among 28 isolates of H3N2 virus, 3 strains were obtained outside the influenza season. Nucleotide sequences of the haemagglutinin (HA) genes of these isolates along with those from domestic patients were analysed in order to determine the influence of imported influenza viruses by travellers on epidemics in Japan. From the phylogenetic and chronological aspects, the possibility was suggested in one case in 1997/8 and two in the 1998/9 season that imported virus by travellers may have influenced the domestic influenza epidemics.  相似文献   

11.
目的了解2009年7月以来杭州地区甲型H1N1流感病毒神经氨酸酶基因(NA)的变异情况,分析其遗传进化特征。方法在流行季节采集发热病人咽拭子样本,经病毒分离培养及亚型鉴定后,用特异性引物扩增NA基因,测序并分析其遗传进化特征。结果 2009年7月至2011年3月,共检测监测样本1 898份,流感病毒阳性率为37.9%。期间经历了两次甲型H1N1流感病毒流行高峰。各时间段分离株间NA基因高度同源,序列相似性97.9%~100%,但2010/2011年流行株在进化树上分为两支,且与2009/2010年流行株遗传距离相对较远。进一步分析后发现,虽然耐药位点、酶活性位点及其附近氨基酸相对保守,但多个氨基酸变异发生于抗原决定簇上,且增加了NA蛋白茎部第42位糖基化位点。结论结果预示着酶抑制剂类药物对预防和治疗甲型H1N1流感病毒仍将有效,但在开发新疫苗时应该尽可能的避开已经发生漂变的抗原决定簇。  相似文献   

12.
符文华  王冰  白杉 《职业与健康》2012,28(10):1176-1179
目的分析甲型H1N1流行性感冒(流感)大流行前后沈阳流感病原学流行特征。方法采集2005年10月—2011年3月连续5.5 a哨点医院的流感样病例咽拭子,进行流感病毒的分离和分型鉴定,并按甲型H1N1流感流行前后划分为3个时期。结果 3个时期共采集流感样病例咽拭子样本6 808份,分离到流感病毒861株,总分离率为12.6%;流行前、流行中和流行后3个时期流感病毒检出率分别为13.8%、18.2%和5.2%。男性和女性流感病毒感染率差异无统计学意义,不同时期流感样病例的年龄构成差异有统计学意义。新的甲型H1N1流感侵袭的人群主要是低年龄组人群,新的甲型H1N1流行期间和流行后,流感季节高峰发生前移。不同年度流感流行毒株型别不同,前1年监测周期的优势株在下1个周期中可被其他型别抑制甚至取代。结论甲型H1N1流感流行后沈阳的流感流行特征已发生了部分改变,这种改变是否持续存在还有待持续监测来得以验证,在2011—2012年监测周期中应密切关注A3(H3N2)亚型和新的甲型H1N1亚型流感毒株的活动情况。  相似文献   

13.
广东省流行性感冒病原学监测结果分析   总被引:12,自引:0,他引:12  
目的分析广东省流行性感冒(流感)流行特点和流感病毒优势株的情况。方法采集2001年1月~2004年6月内科或儿科门诊就诊的流感样病人(发病3d内)的咽拭子或咽漱液标本,流感病毒分离采用鸡胚和狗肾传代细胞(Madin-Darby CanineKidney,MDcK);流感病毒亚型鉴定采用血凝抑制试验。结果共采集流感样病例咽拭子或咽漱液样本17313份,流感病毒阳性1508株,分离率为8.71%,其中A1型(HIN1)92株、A3型(H3N2)1138株、B型278株,分别占总分离数的6.1%,75.5%,18.4%。3种毒株在5~15岁组分布最多,占44.7%;2001年优势流行株为B型,2002~2004年优势流行株均为A3(H3N2);流感病毒分离高峰为每年3~7月,2003年和2004年上半年未分离到A1(H1N1)亚型流感病毒;A3(H3N2)型流感病毒除主要在流行季节分离外,在非流行高峰期也均有一定数量分离;B型流感病毒则主要集中分离于冬季(11、12、1月份)。结论感染人群中流感毒株A1(H1N1)、A3(H3N2)、B型交替出现,A3(H3N2)亚型活动较强。应密切注意A3型(H3N2)流行株的活动情况,加强监测,及时发现流感病毒新变异株。  相似文献   

14.
The pandemic A/H1N1 influenza viruses emerged in both Mexico and the United States in March 2009, and were transmitted efficiently in the human population. They were transmitted occasionally from humans to other mammals including pigs, dogs and cats. In this study, we report the isolation and genetic analysis of novel viruses in pigs in China. These viruses were related phylogenetically to the pandemic 2009 H1N1 influenza viruses isolated from humans and pigs, which indicates that the pandemic virus is currently circulating in swine populations, and this hypothesis was further supported by serological surveillance of pig sera collected within the same period. Furthermore, we isolated another two H1N1 viruses belonging to the lineages of classical swine H1N1 virus and avian-like swine H1N1 virus, respectively. Multiple genetic lineages of H1N1 viruses are co-circulating in the swine population, which highlights the importance of intensive surveillance for swine influenza in China.  相似文献   

15.
目的:对2010年北京市某两区发生的儿童甲3亚型流行性感冒暴发疫情进行病原学研究。方法:荧光RT-PCR方法检测疫情患者标本,对核酸阳性者进行病毒培养;对毒株血凝素HA1基因进行序列比对和进化分析。结果:两起疫情标本甲3亚型流感病毒核酸阳性检出共14件;毒株共分离到6株。各疫情毒株同源性序列为100%;两起疫情的HA1序列同源性为97.3%。与疫苗株A/Perth/16/2009(H3N2)比较,其中疫情2分离株共有8个氨基酸(aa)变异。结论:两起疫情均为甲3亚型(H3N2)流感所致;同一起疫情均由同一病毒株传播所致;两起疫情甲3亚型(H3N2)毒株的血凝素HA1区存在不同程度的aa变异。  相似文献   

16.
目的分析2010-2012年长沙市流感流行情况,并探讨B型流感病毒分离株血凝素(HA)和神经氨酸酶(NA)基因的分子流行病学特征。方法采集长沙市流感网络监测哨点医院及暴发点流感样病例(ILI)咽拭子样本,狗肾传代细胞(MDCK)进行病毒分离,血凝和血凝抑制实验进行型别和亚型鉴定;提取B型流感病毒核酸,一步法RT-PCR扩增HA和NA基因片段,双向测定扩增产物核苷酸序列,对基因序列和氨基酸序列进行分析。结果 2010-2012年间,长沙市甲型H1N1、H3N2、B型流感交替流行,流行高峰为冬春季。3年间共分离到B型流感病毒78株,其中79.5%为Victoria系,20.5%为Yamagata系,以Victoria系为主。与WHO推荐疫苗株B/Brisbane/60/2008相比,HA基因的同源性在87.2%~98.4%之间,NA基因的同源性在94.5%~97.5%之间。氨基酸序列分析发现,与疫苗株相比,HA蛋白主要存在单个氨基酸的突变;种系进化分析发现,所有毒株HA和NA蛋白位于相应的谱系内,无抗原重排发生。结论 2010-2012年间长沙市甲型H1N1、H3N2、B型流感交替流行,B型流感病毒HA基因出现抗原漂移,但无重组毒株的出现。  相似文献   

17.
目的 了解宁夏2018-2019流感监测年度流感病毒病原学检测情况,分析甲型H1N1流感病毒血凝素(HA)基因特征。方法 采用real time RT-PCR方法对流感监测哨点医院采集的流感样病例(ILI)标本进行核酸检测;对阳性标本进行毒株分离;提取甲型H1N1毒株的RNA,采用RT-PCR方法扩增HA片段并测序,利用生物信息软件对测序结果进行比对分析。结果 宁夏流感网络实验室检测咽拭子标本共5214份,核酸检测阳性数为760份,其中甲型H1N1阳性数为485份,占总阳性数的63.82%,分离出甲型H1N1毒株 161株。宁夏分离毒株与疫苗株A/Califaoria/07/2009不在同一进化分支,同源性为92.6%~96.3%;与疫苗株A/Michigan/45/2015(H1N1)为同一进化分支,同源性为96.6%~98.1%。与疫苗株A/Califaoria/07/2009比较,抗原位点、受体结合位点及其他位点均有变异,除毒株 A/Ningxia_Xixia/SWL1176/2019(H1N1)第222位氨基酸发生D222G变异外,其他甲型H1N1流感毒株均未发生D222G变异。所有毒株增加糖基化位点162NQT,个别毒株糖基化位点增加2~3个。结论 宁夏2018 -2019年度流感优势毒株为甲型H1N1毒株。序列分析表明甲型H1N1病毒发生了不同程度的变异,在抗原特异性、毒力和感染性上有可能已经发生变化,需要及时更换疫苗株成分。  相似文献   

18.
OBJECTIVE: Due to the lack of correlation from 1994 to 1997 between the A H3N2 component of the influenza vaccine recommended for this period and the circulating viruses in Argentina, we decided to study the antigenic and genomic relationships of the 1998 A H3N2 Argentine circulating strains with the corresponding vaccine component for that year as recommended by the World Health Organization (WHO). METHODS: We selected 18 influenza A H3N2 strains isolated in Argentina during 1998 to carry out an antigenic and genomic study of their hemagglutinin (HA) and neuraminidase (NA) proteins. For the genomic study we added 3 isolates from Uruguay. We compared the Argentine and Uruguayan strains with available reference strains. RESULTS: We found that all 18 strains from Argentina were similar to the A/Sydney/5/97 (H3N2) strain, as opposed to the A/Wuhan/359/95 (H3N2) strain, which was the vaccine component. This result was confirmed by the genomic study. CONCLUSIONS: The approach that we applied in Argentina has improved the quality and quantity of information about influenza in the country. This type of work should be encouraged in other countries in order to help choose the most appropriate vaccine components each year and provide individuals with the best possible protection against influenza.  相似文献   

19.
分子生物学方法在儿童流行性感冒监测中的应用   总被引:3,自引:1,他引:2  
目的:建立一种能够快速,特异,有效地直接从临床标本中检测流行感冒(流感)病毒并进行病毒型和亚型鉴别的方法,同时了解近年来北京地区A3型流感病毒因凝素重链(HA1)区基因的变异特点。方法:根据编码A、B型流感病毒膜蛋白M基因的核苷酸序列设计两对引物,用于同一逆转录(RT)-PCR反应中(多重RT-PCR),根据A、B型毒株扩增产物的大小(分别为506bp和261bp)鉴别A、B型流感病毒,另根据编码A1和A3型流感病毒糖蛋白HA基因的核苷酸序列设计两对引物,与B型M基因基因引物用同一RT-PCR反应中,可区分A1、A3或B型病毒HA基因和B型病毒M基因又设计了3对引物作为外侧引物,进行巢式-PCR反应。根据第二次PCR的扩增产物在1.2%的琼脂糖凝胶电泳中的大小即可确定型别。经PCR扩增1996-2002年间分离的北京地区A3型流感病毒株HA1区基因,测序并进行序列分析,结果:用上述多重RT-PCR鉴定流感病毒分离株156株,与血凝抑制试验阳性符合率为100%,用多重巢式-PCR检测92份已经病毒分离确定为流感的儿科临床呼吸道标本,与病毒分离和血凝抑制试验阳性符合率分别为76.9%(A1型)、57.1%(A3型)、86.5%(B型),对5株1996-2002年间A3间分离株HA1区基因序列分析结果显示,不同年份的分离株HA1区基因具有较高的核苷酸和氨基酸同源性,而且年份越近的毒株之间同生越测。为流感监测提供更加快速的新方法,北京地区A3型流感毒分离株HA1区基因持续不断地发生点突变,糖基化位点不断增多,可能是导致其抗原漂移的原因。  相似文献   

20.
目的 了解镇江地区甲型H1N1流感病毒流行和变异特点。方法 收集2014-2016年镇江地区哨点医院流感样病例标本,进行核酸检测和病毒分离。在病毒流行期按月随机抽取13株甲型H1N1毒株,设计特异性的引物扩增血凝素(hemagglutinin,HA)、神经氨酸酶(neuraminidase,NA)基因,进行测序并分析其遗传进化特征。结果 13株分离株与疫苗株A/California/07/2009的HA基因核苷酸和氨基酸的同源性分别为97.3%~100.0%和96.6%~100.0%;NA基因核苷酸和氨基酸同源性分别为95.6%~97.5%和93.8%~96.6%。系统进化分析表明,13株病毒HA和NA基因分属于不同的进化谱系。分子特征表现为12株HA氨基酸序列均发生了抗原位点Sa区K173Q突变,1株同时发生了Sa区K181E突变;3株还发生了Ca1区V183I突变。受体结合位点和糖基化位点的氨基酸序列均未发生变异。结论 2014-2016年镇江地区甲型H1N1流感病毒与疫苗株相比,其HA、NA基因出现了一定程度的变异,但是抗原性并未发生改变,需进一步加强监测。  相似文献   

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