2021年银川市4株人呼吸道合胞病毒全基因组基因特征

 目的 分析2021年银川市人呼吸道合胞病毒(HRSV)全基因组的基因特征。 方法 收集2021年银川市2所流感监测哨点医院的流感样标本共1 100份,选择发热并伴有严重呼吸道感染症状的标本共669份,通过实时荧光定量逆转录聚合酶链反应(RT-PCR)法筛选出HRSV阳性标本,对其中符合测序要求的HRSV阳性标本进行全基因组测序。在GenBank数据库下载HRSV全球各基因型别代表株,使用MEGA7软件进行序列比对、亲缘关系进化树构建和遗传距离的计算分析,使用DNAStar软件中MegAlign分析HRSV测序株序列核苷酸(氨基酸)的同源性和氨基酸变异。 结果 669份全年龄段的鼻咽拭子标本,HRSV阳性标本60份,阳性率为8.97％。其中有4份HRSV阳性标本全基因组测序成功,亲缘关系进化树显示,2株HRSVA为ON1型,2株HRSVB为BA9型。2条HRSVA全长序列均与MN306029.1/USA/2019/ON1全长序列的核苷酸(氨基酸)同源性最高,2条HRSVB G蛋白基因序列均与MW160815.1/Australia/2017/BA9的G蛋白基因序列核苷酸(氨基酸)同源性最高。氨基酸变异分析结果表明,2条HRSVA在第113、131、178、257、258、266位均发生了突变,分别为T113I、V131D、N178G、E257K、H258Q、H266L,YinChuanRSVA-01在第134位发生K突变为R(K134R),2条HRSVB在第159位和第160位均出现氨基酸缺失,在G蛋白第二高变区的第254、270、276、290位均发生T突变为I(T254I、T270I、T276I、T290I)。 结论 2021年银川市HRSV出现了RSVA基因型和RSVB基因型的共同流行,以及多个氨基酸位点的变异。     

Rapid replacement of human respiratory syncytial virus A with the ON1 genotype having 72 nucleotide duplication in G gene

Human respiratory syncytial virus (HRSV) is the main cause of severe respiratory illness in young children and elderly people. We investigated the genetic characteristics of the circulating HRSV subgroup A (HRSV-A) to determine the distribution of genotype ON1, which has a 72-nucleotide duplication in attachment G gene. We obtained 456 HRSV-A positive samples between October 2008 and February 2013, which were subjected to sequence analysis. The first ON1 genotype was discovered in August 2011 and 273 samples were identified as ON1 up to February 2013. The prevalence of the ON1 genotype increased rapidly from 17.4% in 2011–2012 to 94.6% in 2012–2013. The mean evolutionary rate of G protein was calculated as 3.275 × 10⁻³ nucleotide substitution/site/year and several positively selected sites for amino acid substitutions were located in the predicted epitope region. This basic and important information may facilitate a better understanding of HRSV epidemiology and evolution.     

Genetic variation of human respiratory syncytial virus among children with fever and respiratory symptoms in Shanghai,China, from 2009 to 2012

Human respiratory syncytial virus (HRSV) of genus Pneumovirus is one of the most common pathogens causing severe acute lower respiratory tract infection in infants and children. No information on the genotype distribution of HRSV is available in East China (e.g. Shanghai). From August 2009 to December 2012, 2407 nasopharyngeal swabs were collected from outpatient children with fever and respiratory symptoms in Shanghai. HRSV infection was determined using a multiplex RT-PCR assay. The second hypervariable region (HVR2) of G protein gene of HRSV was amplified and sequenced from HRSV positive samples. Genotypes were characterized by phylogenetic analyses. Of 2407 nasopharyngeal samples, 184 (7.6%) were tested as HRSV positive. From 160 positive subjects with sufficient nasopharyngeal samples, 69 HVR2 sequences were obtained by RT-PCR and sequencing. Three HRSV epidemic seasons were observed from August 2009 to December 2012, and an extreme outbreak of HRSV occurred in the 2009–2010 epidemic season. A genotype shift of predominant HRSV strains from B group in the 2009–2010 epidemic season to group A in the subsequent epidemic seasons was observed. Ten HRSV genotypes, including four group A genotypes NA1, NA3, NA4, and ON1, and six group B genotypes BA9, BA10, SAB4, CB1, BAc, and BA?, were detected in Shanghai. Seven genotypes (NA1, BA9–10, SAB4, CB1, BAc and BA?) were found in the 2009–2010 epidemic season. The co-circulation of multiple genotypes was associated with the extreme outbreak of HRSV among children with fever and respiratory symptoms in the 2009–2010 epidemic season.     

Genotype circulation pattern of human respiratory syncytial virus in Iran

In order to have information on the molecular epidemiology and genetic circulation pattern of human respiratory syncytial virus (HRSV) in Iran, we studied the genetic variability of both group A and B HRSV strains during seven consecutive years by sequencing the hypervariable C-terminal domain of G protein. A total of 485 children <2 years of age who were negative for influenza viruses, screened for the presence of HRSV in this research. HRSV was detected in 94 (19.38%) of the samples using nested RT-PCR. Group A viruses were isolated during each year, while group B viruses were isolated during 2009 and 2013. Phylogenetic analysis showed that all HRSV group A viruses belonged to three genotypes: GA1, GA2, GA5 and the group B viruses were in BA genotype.     

Molecular evolution of the fusion protein gene in human respiratory syncytial virus subgroup A

We studied the molecular evolution of the fusion protein (F) gene in the human respiratory syncytial virus subgroup A (HRSV-A). We performed time-scaled phylogenetic analyses using the Bayesian Markov chain Monte Carlo (MCMC) method. We also conducted genetic distance (p-distance), positive/negative selection, and Bayesian skyline plot analyses. Furthermore, we mapped the amino acid substitutions of the protein. The MCMC-constructed tree indicated that the HRSV F gene diverged from the bovine RSV (BRSV) gene approximately 550 years ago and had a relatively low substitution rate (7.59 × 10^− 4 substitutions/site/year). Moreover, a common ancestor of HRSV-A and -B diverged approximately 280 years ago, which has since formed four distinct clusters. The present HRSV-A strains were assigned six genotypes based on F gene sequences and attachment glycoprotein gene sequences. The present strains exhibited high F gene sequence similarity values and low genetic divergence. No positive selection sites were identified; however, 50 negative selection sites were identified. F protein amino acid substitutions at 17 sites were distributed in the F protein. The effective population size of the gene has remained relatively constant, but the population size of the prevalent genotype (GA2) has increased in the last 10 years. These results suggest that the HRSV-A F gene has evolved independently and formed some genotypes.     

2013-2017年广州市住院儿童呼吸道合胞病毒流行特征及分子生物学分析

目的分析广州人呼吸道合胞病毒(HRSV)的流行特征及遗传变异特征。方法2013—2017年在广州两家哨点医院(广东省妇幼保健院和中山大学孙逸仙纪念医院)采集0~6岁急性呼吸道感染住院儿童的鼻咽拭子作为标本。采用逆转录PCR和巢式PCR方法进行HRSV的检测和分型,通过MEGA 6.0、NetNGlyc 1.0等软件对HRSV序列进行基因亲缘性分析以及N?糖基化位点的预测。结果共收集鼻咽试子标本1225份,其中男性783份,女性442份;月龄M(P25,P75)为8(3,24)月。共检出HRSV儿童感染者209例(17.06%),其中HRSV?A为117例(55.98%),HRSV?B为92例(44.02%)。2岁以下儿童HRSV检出率为18.83%(196例),占总阳性标本的93.78%。209例检出HRSV儿童中,32例(15.31%)还感染了至少1种其他呼吸道病毒。进化树分析显示,62株HRSV?A属于ON1基因型,2株属于NA1基因型,53株HRSV?B全部属于BA基因型。G蛋白的第二高变区在氨基酸替换,终止密码子的使用以及糖基化位点方面均具有多态性。结论广州2岁以下儿童是感染HRSV的高发人群,ON1为HRSV?A主导基因型,BA为HRSV?B的主导基因型。多样化的氨基酸替换,某些糖基化位点的缺失与插入,体现了G蛋白作为主要保护性抗原的多样性。     

Comparison of antibodies directed against human respiratory syncytial virus antigens present in two commercial preparations of human immunoglobulins with different neutralizing activities

Antibodies directed against human respiratory syncytial virus (HRSV) from two commercial preparations of human immunoglobulins (Igs) were compared. One of the Ig preparations (RespiGam) was obtained from blood samples selected for high titres of anti-HRSV neutralizing antibodies. The other preparation (Flebogamma) was obtained from unselected blood donations. RespiGam and Flebogamma had very similar anti-HRSV ELISA titres, but RespiGam neutralized virus infectivity 8-10 times more efficiently than Flebogamma. The same behaviour was observed when purified antibodies from RespiGam and Flebogamma, specific for either the fusion (F) or the attachment (G) glycoprotein, were compared. To gain further information about differences in neutralization between these two Ig preparations, antibodies recognizing certain F and G protein fragments or peptides were purified and their neutralizing activities were compared. In general, antibodies purified from RespiGam showed higher neutralizing activity that those purified from Flebogamma, but those differences were higher with antibodies specific for certain protein segments than for others. Some of the protein regions recognized by human neutralizing antibodies were mapped outside antigenic sites identified previously with panels of murine monoclonal antibodies. These results offer the possibility of searching for new neutralizing antibodies that could be used to study the molecular basis of neutralization and to prevent HRSV infections.     

13株中国麻疹野病毒血溶素蛋白基因特征分析

目的分析1999～2003年中国（未包括香港、澳门特别行政区和台湾地区,下同）流行的H,基因型麻疹野病毒代表株血溶素蛋白（Fusion Protein,F）基因的分子特征。方法从1999～2003年中国分离的麻疹野病毒株中,选取13株H1基因型代表株,包括H1a基因亚型麻疹野病毒8株和H1b基因亚型麻疹野病毒5株。使用逆转录一聚合酶链反应扩增病毒的F基因全长,并进行核苷酸序列测定,同时与中国疫苗株及其它国家的A、D2、D6、D7、E基因型参考株进行序列比对及基因亲缘性关系分析。结果1999～2003年13株中国流行麻疹野病毒代表株中,核苷酸同源性为98．1％～99．0％,氨基酸同源性为98．1％～100．0％,与中国疫苗株沪191相比,其核苷酸同源性为95．7％～96．2％,氨基酸同源性为96．9％～97．2％;而与其它基因型相比,核苷酸同源性为94．4％～96．2％。F基因3个糖基化位点（第32、64、70位氨基酸）、对病毒融合功能起着重要作用的第112位精氨酸、第195位亮氨酸未发生改变。结论1999～2003年中国流行的H1基因型麻疹野病毒的F基因无明显差异,F蛋白上已知的重要功能位点未发生变异,推测中国流行的麻疹野病毒F蛋白的结构和功能未发生较大改变,但由于F蛋白是重要的功能蛋白,对F蛋白核苷酸和氨基酸的变异规律进行常规监测,对了解麻疹野病毒流行特点及其遗传变异规律具有重要意义。     

人呼吸道合胞病毒分离株基质蛋白基因特征分析

目的 阐明人呼吸道合胞病毒（Human Respiratory Syncytial Virus,HRSV）A、B亚型病毒分离株的基质蛋白（Matrix Protein,M）编码基因特征。方法采用逆转录-聚合酶链反应（Reverse Transciption-Polymerase Chain Reaction,RT-PCR）对北京市2004年分离的HRSV代表株（A、B亚型各2株）进行M基因全长序列扩增、测序,并和基因库下载的所有HRSV的M基因全长序列进行对比和分析。结果2株HRsVA亚型分离株与A亚型Long株（标准株）M基因之间的核苷酸和氨基酸差异,分别为24、26个（3．1％～3．4％）和1个（0．39％）。2株HRSVB亚型分离株与B亚型参考株9320的M基因之间的核苷酸和氨基酸差异,分别为16、19个（2．1％、2．46％）和2、1个（0．78％、0．39％）。所测定的北京市4株HRSV分离株和从基因库下载的M基因全序列之间的核苷酸和氨基酸差异,分别为24～119个（3．1％～15。4％）和1～21个（0．39％～8．2％）。结论M基因在HRSV基因组中是较为保守的基因之一,A或B亚型的型内差异相对较小;但A、B亚型之间有较大差异,变异平均分配于整个M基因中。M基因的这些基因特征使其可以作为A、B亚型基因检测分型诊断的候选靶基因。该研究为HRsV基因快速诊断试剂的研制提供了基因信息学数据。     

Evolutionary analysis of human respiratory syncytial virus collected in Myanmar between 2015 and 2018

We studied genetic variation in the second hypervariable region (HVR) of the G gene of human respiratory syncytial virus (HRSV) from 1701 nasal swab samples collected from outpatients with acute respiratory infections at two general hospitals in the cities Yangon and Pyinmana in Myanmar from 2015 to 2018. HRSV genotypes were characterized using phylogenetic trees constructed using the maximum likelihood method. Time-scale phylogenetic tree analyses were performed using the Bayesian Markov chain Monte Carlo method. In total, 244 (14.3%) samples were HRSV-positive and were classified as HRSV-A (n = 84, 34.4%), HRSV-B (n = 158, 64.8%), and co-detection of HRSV-A/HRSV-B (n = 2, 0.8%). HRSV epidemics occurred seasonally between July (1.9%, 15/785) and August (10.5%, 108/1028), with peak infections in September (35.8%, 149/416) and October (58.2%, 89/153). HRSV infection rate was higher in children ≥1 year of age than in those <1 year of age (70.5% vs. 29.5%). The most common HRSV symptoms in children were cough (80%–90%) and rhinorrhea (70%–100%). The predominant genotypes were ON1for HRSV-A (78%) and BA9 for HRSV-B (64%). Time to the most recent common ancestor was 2014 (95% highest posterior density [HPD], 2012–2015) for HRSV-A ON1 and 2009 (95% HPD, 2004–2012) for HRSV-B BA9. The mean evolutionary rate (substitutions/site/year) for HRSV-B (2.12 × 10⁻², 95% HPD, 8.53 × 10⁻³–3.63 × 10⁻²) was slightly higher than that for HRSV-A (1.39 × 10⁻², 95% HPD, 6.03 × 10⁻³–2.12 × 10⁻²). The estimated effective population size (diversity) for HRSV-A increased from 2015 to 2016 and declined in mid-2018, whereas HRSV-B diversity was constant in 2015 and 2016 and increased in mid-2017. In conclusion, the dominant HRSV-A and HRSV-B genotypes in Myanmar were ON1 and BA9, respectively, between 2015 and 2018. HRSV-B evolved slightly faster than HRSV-A and exhibited unique phylogenetic characteristics.     

High genetic diversity and superinfection by Anaplasma marginale strains in naturally infected Angus beef cattle during a clinical anaplasmosis outbreak in southeastern Brazil

Anaplasma marginale is an obligate intracellular Gram-negative bacterium that is parasitic to erythrocytes and is the main agent of bovine anaplasmosis. This disease causes severe anemia and reduces weight gain and milk production, thus giving rise to major economic losses relating to livestock worldwide. The genetic diversity of this bacterium has been characterized based on sequences of major surface proteins (MSPs), especially MSP1α. This has enabled identification of several geographical strains, according to different amino acid sequences. The aim of this study was to investigate the genetic diversity of A. marginale in naturally infected Angus beef cattle during a disease outbreak in southeastern Brazil. Four blood samples were collected over a four-month period from each of 20 animals on a cattle farm in Itú, São Paulo, Brazil. Serum samples were subjected to indirect ELISA to detect anti-A. marginale IgG antibodies. The 80 whole-blood samples obtained were subjected to DNA extraction, quantitative real-time PCR (qPCR) for the msp1β gene, semi-nested PCR (snPCR) for the msp1α gene, cloning of the target fragment and sequencing using the Sanger method. The sequences obtained were analyzed for genetic diversity using the RepeatAnalyzer software. Both iELISA tests, using recombinant MSP5 and the Anaplasma antibody test kit (VMRD), revealed high seroprevalence: 91.25% and 97.5%, respectively. In qPCR, 100% of the samples were positive, with between 10³ and 10⁷ DNA copies/μL. In the snPCR based on the msp1α gene, 57.5% (46/80) of the samples were positive. Microsatellite analysis on the 36 sequences obtained showed the presence of genotypes H (58.3%), F (25%), E (19.4%), C (2.7%) and G (2.7%). The RepeatAnalyzer software identified 36 strains in the study region, among which some had not previously been described in the literature (13 27 13 27 13 F; 16 FF; τ 27; 63 29 104 29; LJ1 13 LJ1 13; 16 F 17; 16 F 91). High genetic diversity of A. marginale bacteria was found on this farm in Itú, São Paulo.     

Genotyping of Echinococcus granulosus Isolates from Human Clinical Samples Based on Sequencing of Mitochondrial Genes in Iran,Tehran

Background

The present study was aimed to investigate molecular diversity of Echinococcus granulosus isolates collected from human clinical samples using two mitochondrial genes cox1 and nad1 in Iran.

Methods

Forty seven human hydatid cysts were collected through surgery from two hospitals in Tehran during 2010-2012. To determine the fertility of protoscoleces, the cyst fluids were subjected to morphological microscopic examinations. Protoscoleces were removed from each cyst and their total genomic DNAs were extracted. PCR was performed to amplify fragments of 450 and 400 base pair (bp) for cox1 and nad1 genes, respectively. Genotype diversity and sequence variation of the strains were studied by bioinformatics software and in comparison with those mtDNA sequences already deposited in GenBank.

Results

Sixteen, (53.3%), 13 (43.3%), and 1 (3.3%) samples were related to lung, liver, and spleen, respectively. The remained 17 unfertile samples were excluded from the study. From the 29 isolates, 86.7% (n=26) and 10% (n=3) were related to G1, and G3 genotypes, respectively. The sole isolate with G6 genotype was obtained from lung sample. Analysis of concatenated sequences of cox1+nad1 indicated the presence of 11 haplotypes among our strains that were related to genotypes G1 (n=9), G3 (n=1) and G6 (n=1).

Conclusion

In consistent to other reports from Iran, genotypes G1, G3, and G6 were observed in our human isolates. The rate of G3 genotype was however higher than other studies implying that human can be considered as a new appropriate host for G3 genotype. Further studies with more sample size from different geographic areas of Iran are needed for E. granulosus mapping.     

Diversity of VP7 genes of G1 rotaviruses isolated in Iran, 2009–2013

Genotype G1 of rotaviruses (RVs) is the most prevalent strain in human RV infections around the world. The present study evaluated genetic variations in the VP7 gene of RV G1 genotype isolates from Iran. Genetic and phylogenetic analyses indicated that RV strains from Iran clustered with G1 lineages IA, IC, and IIC, showing highest average of similarity versus reference sequences of the G1 lineages I and II. This study highlights the genetic pattern of G1 RV on the basis of distinct lineages and sublineages and indicates the importance of continuous monitoring on genetic variation and evolution pattern of G1 RV strains across the Iranian population for the final aim of RV vaccine introduction.     

Proposal for Human Respiratory Syncytial Virus Nomenclature below the Species Level

Human respiratory syncytial virus (HRSV) is the leading viral cause of serious pediatric respiratory disease, and lifelong reinfections are common. Its 2 major subgroups, A and B, exhibit some antigenic variability, enabling HRSV to circulate annually. Globally, research has increased the number of HRSV genomic sequences available. To ensure accurate molecular epidemiology analyses, we propose a uniform nomenclature for HRSV-positive samples and isolates, and HRSV sequences, namely: HRSV/subgroup identifier/geographic identifier/unique sequence identifier/year of sampling. We also propose a template for submitting associated metadata. Universal nomenclature would help researchers retrieve and analyze sequence data to better understand the evolution of this virus.     

中国2006年麻疹野病毒血溶素基因特征分析

目的研究中国2006年流行的麻疹野病毒代表株血溶素蛋白（Fusion Protein,F）基因的分子特征,与其他年份毒株相比分析F基因的变异规律。方法从2006年各省（自治区、直辖市）送检的麻疹野病毒中选取9株代表株,使用逆转录-聚合酶链反应扩增病毒的F基因全长,并进行核苷酸序列测定,同时与中国疫苗株及其他年份毒株进行序列比对及基因亲缘性关系分析。结果2006年9株中国流行麻疹野病毒代表株,其核苷酸同源性为98．5％-99．8％,氨基酸同源性为99％-100％;与中国疫苗株沪。相比,其核苷酸同源性为95．4％-96．2％,氨基酸同源性为96．7％～97．2％;与1999～2003年代表株相比,其核苷酸同源性为97．9％-99．7％,氨基酸同源性为98．1％-100％。F基因3个糖基化位点（第32、64、70位氨基酸）、对病毒整合功能起着重要作用的第112位精氨酸（Arg）、第195位亮氨酸（Leu）未发生改变。结论中国2006年流行的麻疹病毒F基因变异不大,与其他年份毒株相比变异也不明显。F蛋白的重要功能位点未发生变异,F蛋白的变异与2006年麻疹流行无相关性。但由于F蛋白对病毒感染细胞和促使机体产生抗体具有重要意义,对F蛋白变异规律进行常规监测,对了解麻疹病毒变异规律和评价疫苗的保护效果具有重要意义。     

Analysis of the genetic variability of the mumps SH gene in viruses circulating in the UK between 1996 and 2005

Genetic analysis (genotyping) of mumps viruses has been applied to the molecular epidemiology of mumps for over 10 years in the UK. To explore further the variation of mumps strains over time, in total, 965 sequences of the entire SH gene were analysed and compared, including 954 mumps virus strains collected in the UK between 1996 and 2005 were characterised as genotypes G2 (426), G5 (369), J (157) and F (2), which were compared with 11 F sequences found in China. Phylogenetic trees drawn for G2, G5 and J sequences showed that the diversities were greater between the sequences in earlier years (before 2001/2002) than those in later years and could be divided into two clusters within each of the three genotypes over the 10-year period. One transmission of G2, G5 and a J strain was sustained from earlier years with mutations and eventually became predominant strains. Divergences amongst the same genotype or sub-genotype was up to 4.6% for G2, 5.3% for G5 and 4.9% for J. Mutation rates per site per year based on the 316 nt of SH gene were 0.94, 1.3, 0.96 and 1.86 × 10^?2 for G2, G5, J and F respectively. The ratio of d_N/d_S was 0.556, 0.909, 0.357 and 0.811 calculated based on the sequences of G2, G5, J and F respectively. The results revealed that the possible mumps evolution process based on the SH gene was not driven by positive selection during the 10 years between 1996 and 2005.     

Molecular epidemiology and genetic diversity of hepatitis B virus in Mar del Plata city,Argentina

The aim of this work was to describe the current molecular epidemiology and genetic diversity of HBV in Mar del Plata, an important Argentinean touristic city. The phylogenetic analysis of 29 HBV DNA positive serum samples showed that F1b was the predominant subgenotype (sgt, 62.1%), followed by sgt A2 (13.8%) and sgt F4, gt D and gt G (6.9% each). Among anti-HBc IgM positive samples, 75.0% were sgt F1b, followed by sgt F4 (12.5%), sgt A2 (6.25%) and sgt D (6.25%). Three recombinant full length genomes were found: two G/F1b (some of the first gt G detected in Argentina) and one F4/D2. The circulation of clinical important mutations in the city was described. Mutations at the HBsAg were detected in 34.5% of the analyzed samples, associated with laboratory diagnosis and antiviral treatment failures, immune escape and hepatocellular carcinoma. Most of the samples presented wild type BCP/PC sequences. Coalescence analysis for the most prevalent sgt F1b estimated that the diversification mainly occured during mid '90s and the tMRCA was estimated in 1987. Finally, the high presence of the autochthonous sgt F1b, associated with the anti-HBc IgM positive infection and its present-day diversification process, shows the strong impact of internal human migratory movements into the current population of Mar del Plata.     

Molecular Differentiation of Fasciola Species and Characterization of Genetic Diversity of F. gigantica Using NADH Dehydrogenase I (ND1) Gene in the Endemic Areas of Iran

Background:

Fasciola hepatica and F. gigantica are the causative agents of fasciolosis in domestic animals and humans. Based on the morphometric criteria, differential diagnosis between them is problematic. In addition, intermediate forms of Fasciola have been found in Iran, which makes the differentiation more difficult. The aim of the present study was to provide molecular evidence for the existence of F. gigantica in Iran using sequencing analysis of ND1 and PCR-RFLP analysis of ITS2 regions and to study the intraspecies variations of F. gigantica based on mitochondrial ND1 gene polymorphism.

Methods:

Forty Fasciola spp. samples collected from four distinct provinces (Fars, Khuzestan, Gilan, Khorasan Razavi) in Iran were collected for morphological and molecular characterization. In molecular method, PCR-RFLP analysis of ITS2 using pagI restriction enzyme was used as a screening approach for F. gigantica differentiation. Then mitochondrial DNA sequence variations in the ND1 gene were used for phylogenetic analysis.

Results:

Based on the morphometric criteria and RFLP analysis, 14 parasitic samples were initially identified to be F. gigantica. Phylogenetic results showed that there are at least 10 different genotypes of F. gigantica in Iran, which are different from those existing in the GenBank. Twenty-six points out of 410 base pairs of sequenced ND1 gene in 10 varieties of F. gigantica were diagnosed to be polymorphic. From 26 points of polymorphism, only eight resulted in the post-translational amino acid changes in ND1 gene product structure.

Conclusion:

Data revealed noticeable genetic diversity (up to 4.63%) between different varieties of F. gigantica in Iran.     

杭州地区2009-2010年急性腹泻患者中杯状病毒的检测和分型

目的 了解2009-2010年杭州地区急性腹泻患者中杯状病毒基因型别和分子流行病学特征.方法 收集2009-2010年杭州地区920例急性腹泻患者的粪便标本和流行病学资料,用多重PCR方法进行杯状病毒检测,测定部分标本的阳性扩增片段基因序列,并对序列进行系统进化分析.结果 急性腹泻患者杯状病毒检出率为21.8％(201/920),其中诺如病毒(NV)G Ⅰ型25例、GⅡ型170例、札如病毒(SAV) 11例,4例为NVⅠ型和Ⅱ型混合感染、1例为NVⅡ型和SAV混合感染.NV基因型别包括:GⅠ-1(3株)、G Ⅰ-2(1株)、GⅡ-4/2006b变异株(7株)、GⅡ-2(1株)、GⅡ-7(1株)和GⅡ-4/2008变异株(2株);SAV基因型别包括:G Ⅰ -2(5株)、GⅠ-1(4株)和GⅡ-1(1株).杯状病毒的流行在不同季节、年龄组人群均有发病.结论 杯状病毒是2009-2010年引起杭州地区急性腹泻的主要病原之一,其病原具有病毒多样性和基因型别多样性,NVGⅡ-4/2006b变异株或类似株可能是2009-2010年杭州地区流行的优势株.