山东地区输血后丙肝患者HGV感染状况及HGV部分核酸序列测定

应用ＲＴ－ＰＣＲ法检测庚肝病毒（ＨＧＶ）ＲＮＡ,并对阳性扩增的产物采用ＰＣＲ片段直接克隆法测定。结果从８６例输血后丙型肝炎（ＰＴＨ－Ｃ）患者血清中检出ＨＧＶＲＮＡ阳性者３１例（３６．０％）,其中１例ＨＧＶＮＳ５区部分核苷酸序列与美国原始株和另１例日本株核苷酸同源性比较分别为８６．０％和８４．０％。证实山东地区ＰＴＨ－Ｃ２存在着ＨＧＶ感染和ＨＧＶ／ＨＣＶ混合感染,但是否存在不同亚型或ＨＧＶＲＮＡ阳性     

从长春分离的一株急性散发性戊型肝炎病毒部分核苷酸序列分析

目的分析长春一株急性散发性戊型肝炎病毒（ＨＥＶ）的部分核苷酸序列。方法应用逆转录套式聚合酶链反应法检测ＨＥＶＲＮＡ；用荧光法（ＡｐｐｌｉｅｄＢｉｏｓｙｓｔｅｍｓ）直接测序。结果被检的１４例急性散发性戊型肝炎病人中，５例ＨＥＶＲＮＡ阳性。对其中一例病人的ＨＢＶｃＤＮＡ开放读框２区进行核苷酸序列分析并与墨西哥株（Ｍ）和缅甸株（Ｂ）比较，发现长春散发性ＨＥＶｃ－１８７株与ＨＥＶ（Ｍ）核苷酸和氨基酸序列的同源性分别为７９．９％和９３．１％；与ＨＥＶ（Ｂ）散发株和流行株的同源性分别为９２．３％和９６．２％及９３．９％和９７．７％；与新疆株（ＣＨ１．１）同源性分别为９６．５％和９５．４％。结论长春散发性ＨＥＶｃ－１８７株与新疆株（ＣＨ１．１）一样，可能与ＨＥＶ（Ｂ）为同一亚型。     

粪便中戊型肝炎病毒散发株的核酸序列分析

目的了解粪便中戊型肝炎病毒（ＨＥＶ）散发株的核酸序列变异情况。方法用逆转录－套式－聚合酶链反应（ＲＴ－Ｎｅｓｔｅｄ－ＰＣＲ）检测了８例广州地区散发性戊型肝炎患者粪便中的ＨＥＶＲＮＡ，３例阳性。对阳性ＰＣＲ产物的２３８个碱基进行了基因克隆和核酸序列分析，并与已报道的ＨＥＶ序列进行比较。结果本地区３株ＨＥＶ与缅甸株（Ｂ）、巴基斯坦株（Ｐ）、墨西哥株（Ｍ）、中国新疆株（Ｃｈ１．１）和广州血清株（Ｇ－９）的核苷酸和氨基酸的同源性均值分别为８０．６７％和８８．６０（Ｂ）、８１．２５％和８９．２０％（Ｐ）、７７．４５％和８４．８１％（Ｍ）、８１．２５％和８９．２０％（Ｃ）、９７．８５％和９６．２０％（Ｇ）。结论本组３株ＨＥＶ有一定程度的变异。     

引起细胞病变的甲肝病毒BJ—1株结构蛋白VP4—VP2基因序列的测定

测定我室分离的甲型肝炎病毒ＢＪ－１株结构蛋白ＶＰ４－ＶＰ２基因区的核心苷酸序列。方法从ＢＪ－１感染的Ａ５４９细胞中提取病毒ＲＮＡ模板，通过ＲＴ－ＰＣＲ扩增相应的ｃＤＮＡ片段，测定其核苷酸序列。结果ＢＪ－１株与野型株ＭＢＢ比较，ＶＰ４－ＶＰ２区有４个核苷酸变异，同源性为９９．５％，推论的氨基酸序列有１个氨基酸变异，同源性为９９．６％。与ＨＭ－１７５比较，有３０个核苷酸变异，同源性为９５．９％（７０５     

原发性肝癌患者血清和肝组织中庚型肝炎病毒基因的PCR检测

用逆转录－聚合酶链反应法（ＲＴ－ＰＣＲ）检测了原发性肝癌（ＰＬＣ）患者血清及肝癌和癌旁肝组织中的庚型肝炎病毒（ＨＧＶ）ＲＮＡ，以ＰＣＲ－双脱氧末端终止法测定了ＰＣＲ产物的核苷酸序列。结果显示，血清和肝组织中ＨＧＶＲＮＡ的检出率分别为１９．４％（１３／６７）和２５．７％（９／３５），且ＨＧＶＲＮＡ在肝癌组织呼吕旁肝组织中同时存在，血清和肝组织中ＨＧＶ ＲＮＡ检测结果的符合率为８５％；５′非编码区（５     

慢性非甲—戊型肝炎患者血清中庚型肝炎病毒核酸的检测及部分核苷…

目的 了解庚型肝炎病毒（ＨＧＶ）在慢性非甲－戊型肝炎发病中的作用，并测定其非结构基因５（ＮＳ５）区部分核苷酸序列，方法 以逆转录－套式－聚合酶链反应法检测ＨＧＶＲＮＡ，并对阳性者血清中扩增出的９９４ｂｐ长的ｃＤＮＡ直接测序。结果 ３５例慢性非甲－戊型肝炎患者中１例血清ＨＧＶＲＮＡ阳性（２．９％）。该患者ＨＧＶＮＳ５区部分核苷酸序列与美国株ＰＮＦ２１６１及Ｒ１０２９１的同源性分别为８７．９５％及８９     

上海地区Ⅱ、Ⅲ型丙型肝炎病毒包膜区cDNA序列分析

目的 分析上海地区丙型肝炎病毒（ｈｅｐａｔｉｔｉｓ Ｃ ｖｉｒｕｗ,ＨＣＶ）Ⅱ、Ⅲ型包膜区ｃＤＮＡ序列变异。方法 采用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）扩增ＨＣＶ包膜区Ｅ１、Ｅ２／ＮＳ１片断（１ ００５ｂｐ）,在３７３ＤＮＡ全自动测序仪上进行了序列分析。结果 ＨＣＶ－Ｅ１、Ｅ２／ＮＳ１区核苷酸和氨基酸同源性比较显示：上海株Ⅱ型、Ⅲ型间同源性分别为６２．４８％和５９．１０％;上海株与加内外同型株间     

血清学标志阴性的病毒性肝炎病原学研究

目的 了解血清学杜的病毒性肝炎病原学。方法 应用ＢＨＶ、ＨＣＶ、戊型肝炎病毒（ＨＥＶ）和庚型肝炎病毒（ＨＧＶ）核酸ＰＣＲ检测１０４例血清学标志阴性的病毒性肝炎病人,并用直接测序法对部分ＨＣＶ ＲＮＡ和ＨＥＶ ＲＮＡ逆转录ＰＣＲ产物进行了核苷酸序列测定。结果 １０４例血清学标志阴性的病毒性肝炎病人中,ＨＢＶ ＤＮＡ、ＨＣＶ ＲＮＡ、ＨＥＶ ＲＮＡ和ＨＧＶ ＲＮＡ阳性率分别为２９．８％、３．８％、２２     

散发性戊型肝炎的病原学和血清学研究

目的 研究散发性戊型肝炎（ＨＥ）患者病毒血症,粪便排病毒及抗体消长规律。方法 采用逆转录－套式－聚合酶链反应（ＲＴ－ｎｅｓｔｅｄ－ＰＣＲ）动态检测血清和粪便ＨＥＶＲＮＡ,采用ＨＥＶＯＲＦ２,ＯＲＦ３合成多肽单独和联合ＥＩＡ法动态检测血清抗－Ｈ。结果 血清ＨＥＶＲＮＡ阳性患者占７１％（４４／６２）平均持续时间为病后２０．６天（全程随访３２例）粪便ＨＥＶＲＮＡ阳性标本中有９６．２％（２５／２６）出现在     

戊型肝炎病毒变异株的血清学特征

探讨戊型肝炎病毒（ＨＥＶ）患者血清学特征。方法用逆转录聚合酶链反应法（ＲＴ－ＰＣＲ）检测ＨＥＶＲＮＡ,并对阳性产物进行克隆和测序;对部分ＨＥＶＲＮＡ阳性者进行追踪并检测抗一ＨＥＶ。结果１５例（２３.４％）为ＨＥＶＲＮＡ阳性,其中９例为典型的中国ＨＥＶ株基因序列,６例变异较大,与典型的中国株基因序列的同源性仅为８０％左右。感染原型ＨＥＶ株的４例中,３例于１月后抗一ＨＥＶ阳转;而２例感染变异ＨＥＶ株的患者于１月或３月时,抗－ＨＥＶ仍为阴性。结论现行的抗一ＨＥＶＥＩＡ试剂尚不能诊断变异的ＨＥＶ株。     

Genetic heterogeneity of hepatitis E virus recovered from Japanese patients with acute sporadic hepatitis

The recent discovery of a presumably Japan-indigenous hepatitis E virus (HEV) strain (JRA1) spurred analysis of additional isolates from 7 cases of acute sporadic hepatitis E infection. Comparison of a 326-nucleotide region from open-reading frame 1 indicated that 1, 3, and 3 isolates segregated to genotypes I, III, and IV, respectively. Six patients had not traveled abroad recently. One patient had traveled to Hawaii 1 month before becoming ill, and the nucleotide sequence of the HEV isolate infecting her resembled those of US isolates (89%-91% nucleotide identity). However, the isolate was even more homologous to 2 other Japanese isolates (95%-97% nucleotide identity), suggesting that it is more likely a domestic, rather than an imported, strain. Three genotype IV isolates from Japan also had a higher homology to each other (100% amino acid identity) than to 2 Chinese isolates (97%-98% amino acid identity). These findings suggest that HEV strains of at least 3 different genotypes have already made inroads and are spreading in Japan.     

Molecular Epidemiology and Evolution of Coxsackievirus A9

Nineteen CVA9 isolates were obtained between 2010 and 2019 from six provinces of mainland China, using the HFMD surveillance network established in China. Nucleotide sequencing revealed that the full-length VP1 of 19 CVA9 isolates was 906 bases encoding 302 amino acids. The combination of the thresholds of the phylogenetic tree and nucleotide divergence of different genotypes within the same serotype led to a value of 15–25%, and enabled CVA9 worldwide to be categorized into ten genotypes: A–J. The phylogenetic tree showed that the prototype strain was included in genotype A, and that the B, C, D, E, H, and J genotypes disappeared during virus evolution, whereas the F, I, and G genotypes showed co-circulation. Lineage G was the dominant genotype of CVA9 and included most of the strains from nine countries in Asia, North America, Oceania, and Europe. Most Chinese strains belonged to the G genotype, suggesting that the molecular epidemiology of China is consistent with that observed worldwide. The 165 partial VP1 strains (723 nt) showed a mean substitution rate of 3.27 × 10⁻³ substitution/site/year (95% HPD range 2.93–3.6 × 10⁻³), dating the tMRCA of CVA9 back to approximately 1922 (1911–1932). The spatiotemporal dynamics of CVA9 showed the spread of CVA9 obviously increased in recent years. Most CVA9 isolates originated in USA, but the epidemic areas of CVA9 are now concentrated in the Asia–Pacific region, European countries, and North America. Recombination analysis within the enterovirus B specie (59 serotypes) revealed eight recombination patterns in China at present, CVB4, CVB5, E30, CVB2, E11, HEV106, HEV85, and HEV75. E14, and E6 may act as recombinant donors in multiple regions. Comparison of temperature sensitivity revealed that temperature-insensitive strains have more amino acid substitutions in the RGD motif of the VP1 region, and the sites T283S, V284M, and R288K in the VP1 region may be related to the temperature tolerance of CVA9.     

Swine hepatitis E virus in rural southern China: genetic characterization and experimental infection in rhesus monkeys (Macaca mulatta)

BACKGROUND: In rural areas of southern China, where hepatitis E is endemic, residents generally rear pigs in pigsties near their houses. The study was conducted to assess the possibility that hepatitis E virus (HEV) infections in this region are acquired primarily through contact with swine. METHODS: One hundred twenty swine fecal samples collected from pigsties located in eight rural communities of southern China were tested for HEV RNA. The swine HEV isolates were analyzed genetically and were experimentally inoculated into rhesus monkeys to determine the potential risk of cross-species infection. RESULTS: Twenty-nine of the 120 swine fecal samples were positive for HEV RNA. The nucleotide sequences of these swine HEV strains shared 85%-99% identities with the local human genotype 4 isolates and belonged to two subgroups of genotype 4. Importantly, swine HEV strains representing both subgroups induced hepatitis in rhesus monkeys by inoculation with the virus, evidenced by elevated serum alanine transaminase (ALT), viremia, fecal viral shedding, anti-HEV seroconversion, and liver histopathological changes. CONCLUSIONS: Swine may be the principal reservoir for human HEV infection in rural southern China.     

云南省大理市啮齿动物携带冠状病毒和戊型肝炎病毒的调查

目的 调查云南省大理市啮齿动物冠状病毒(Coronavirus, CoV)和戊型肝炎病毒(Hepatitis E Virus, HEV)感染率。方法 2020年8月～2021年8月在大理市4个乡镇采集啮齿动物样本,采用巢式PCR扩增CoV和HEV的保守序列基因,用生物信息学软件进行同源性与遗传进化分析。结果 共捕获76只啮齿动物属于5属6种,CoV感染动物为褐家鼠(Rattus norvegicus)和黄胸鼠(Rattus tanezumi),感染率分别为40.74%(11/27)、2.38%(1/42);HEV感染动物为褐家鼠和黄胸鼠,感染率分别为14.81%(4/27)、2.38%(1/42)。在银桥镇捕获的2只褐家鼠中同时检测到CoV和HEV,感染率为7.41%(2/27)。基于CoV的部分RNA依赖的RNA酶(RNA-dependent RNA polymerase, RdRp)的434 bp核苷酸片段分析,11株CoV阳性样本均来自褐家鼠,为α-CoV,与来自中国福建黄毛鼠(Rattus losea)RtRl/FJ2015同源性最高,为99.73%～99.74%;1株CoV阳...     

斑点热群立克次体中国分离株HL-93和HLJ-054亲缘关系的研究

目的　通过斑点热群立克次体 (spottedfevergrouprickettsiae ,SFGR)rOmpA基因片段的序列分析对HL 93和HLJ 0 5 4的亲缘关系进行研究。方法　用PCR分段扩增rOmpA基因重复区后 1 9kb片段 ,PCR产物直接测序 ,用WinstarDNA分析软件包进行同源性比较、进化树分析及酶切位点的分析。　结果　HL 93株和HLJ 0 5 4株核苷酸及推定氨基酸的同源性均在 99%以上 ,在树形图上单独聚为一类。常用酶的酶切位点完全一致。结论　HL 93株和HLJ 0 5 4株为同一种SFGR ,建议均命名为黑龙江立克次体 ,但可能存在株间差异性。     

Hepatitis E virus infection among animals in northern India: an unlikely source of human disease

Hepatitis E virus (HEV) is a major cause of acute hepatitis in many developing countries. Based on data from nonendemic regions, an animal reservoir of HEV has been proposed; however, data from HEV-endemic regions are limited. We tested sera from 200 pigs, 98 chickens, 86 goats, 58 sheep and 30 buffaloes for anti-HEV IgG using two different enzyme immunoassays. Specificity of the detected antibodies was confirmed using inhibition assays. Stool specimens from 210 pigs, 94 piglets and 37 sheep were tested for HEV-RNA using nested amplification methods; the polymerase chain reaction products were sequenced and compared with known human and swine HEV sequences. Of the 200 swine sera, 193 and 195, respectively, tested positive in the two assays. All goat sera showed anti-HEV reactivity in both the assays. Inhibition studies confirmed the HEV specificity of antibodies detected in swine and goat sera using both the assays. Sera from sheep, buffalo and chickens also showed high rates of apparent reactivity, but inhibition studies were unable to confirm the specificity of reactions in these species. One faecal specimen showed amplification using Indian swine HEV-specific primers. The genomic sequence of the amplicon from this isolate had only 76-79% nucleotide and 93% amino acid homology with human HEV isolates reported from India and other parts of the world, and most closely resembled swine HEV isolates from other parts of India. Infection with HEV or a related agent is widespread among animals in northern India. However, the swine HEV in India differs genetically from human HEV isolates, indicating that pigs may not play an important role in the spread of human hepatitis E in endemic regions.     

中国12株狂犬病街毒株G基因序列测定与分析

目的了解中国狂犬病毒的流行情况以及街毒株与中国人用、兽用狂犬病疫苗株在G基因核苷酸和氨基酸水平的差异,为有效控制狂犬病疫情提供初步科学依据。方法对12株街毒株G基因进行了全基因测序,与其它36株中国狂犬病街毒株,以及中国的疫苗株和其它国家毒株的G基因序列进行了综合分析。结果序列分析表明来源于中国的50株狂犬病毒均为基因Ⅰ型狂犬病毒;其中具有代表性的6株病毒与我国现在使用的各种疫苗株在G基因的核苷酸和氨基酸水平上均存在不同程度的差异,与我国人用疫苗株CTN同源性较高;进化分析表明,我国主要流行狂犬病毒与泰国、印度尼西亚、马来西亚等东南亚狂犬病毒株处于同一分支。中国毒株之间G基因核苷酸同源性分别≥82.3%;氨基酸同源性分别≥92.1%;中国街毒株与疫苗株相比较核苷酸的同源性为79.3%～94.2%,氨基酸的同源性为87.8%～97.9%。结论我国的狂犬病毒为基因Ⅰ型狂犬病毒,可以明确分为6个进化群。无论在核苷酸还是氨基酸水平上,中国多数街毒株与疫苗株CTN之间的同源性要高于与其它疫苗株。     

狂犬病毒疫苗株CTN-1八代次N和G基因序列测定及分析

目的测定八个代次CTN-1疫苗株核蛋白（N）和糖蛋白（G）基因序列，并与代表性疫苗株和街毒株进行比较。方法用RT-PCR反应扩增八代次CTN-1的N和G基因，获得cDNA，进行序列测定。结果八个代次CTN-1的N和G基因序列与其它疫苗株比较，核苷酸的同源性分别为82．7％～84．5％和86．6％～88．5％，氨基酸同源性分别为88．9％～92．5％和95．3％～98．2％；与在我国分离的街毒株相比，核苷酸的同源性分别为84．2％～95．0％和80．4％～94．3％，氨基酸同源性分别为92．1％～99．6％和88．7％～98．5％。CTN-1株八个代次之间的核苷酸序列几乎完全相同。结论CTN-1株在传代过程中N和G基因变异小，与中国流行的代表性街毒株的同源性高于其它疫苗株。