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Molecular characterization of serotype G2 and G3 human rotavirus strains that have an apparently identical electropherotype of the short RNA pattern

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ARCHIVES OF VIROLOGY 2002年第11期147卷 2187-2195页

作者： Nakagomi, T Gentsch, JR Das, BK Kumar, R Bhan, MK Glass, RI Nakagomi, O Akita Univ Sch Med Dept Microbiol Akita 010 Japan Ctr Dis Control & Prevent Div Viral & Rickettsial Dis Ctr Infect Dis Atlanta GA USA All India Inst Med Sci Dept Pediat Div Gastroenterol & Enter Infect New Delhi India All India Inst Med Sci Dept Microbiol Div Gastroenterol & Enter Infect New Delhi India

The literature is conflicting whether or not rotavirus strains with different G serotype have an identical electropherotype. This is a contentious but an important issue because large parts of molecular epidemiological studies of rotaviruses have been based on the conception that a single strain of rotavirus can be defined by a single electropherotype. Here, we examined in detail by reverse-transcription PCR genotyping, electropherotyping, sequencing, and genogrouping using RNA-RNA hybridization three human rotavirus strains isolated in India that had apparently identical electropherotypes although one strain was typed as P[4], G3 while the other two typed as P[4], G2. These three strains showed an identical electropherotype on 7.5% and 12.5% polyacrylamide gels, but co-electrophoresis on a 10% gel demonstrated that segment 8 of the P[4], G3 strain migrated more slowly than the cognate segment of the P[4], G2 strains. Genogrouping assay and nucleotide sequencing provided evidence for the hypothesis that the P[4], G3 stain was an intergenogroup reassortant in which a P[4], G2 strain of the DS-1 genogroup had acquired the VP7 gene from an yet-unidentified concurrently circulating G3 strain. While electropherotyping remains a valuable asset for molecular epidemiology of rotaviruses, this study underscores the importance of co-electrophoresis under different electrophoretic conditions when pinpointing subtle differences.

关键词：抗原病毒碱基序列衣壳蛋白质类/遗传学电泳聚丙烯酰氨凝胶基因型分子序列数据 RNA 病毒/分析轮状病毒属/分类轮状病毒属/遗传学血清分型人类

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SUBGROUP-I SEROTYPE-3 HUMAN ROTAVIRUS STRAINS WITH LONG RNA PATTERN AS A RESULT OF NATURALLY-OCCURRING REASSORTMENT BETWEEN MEMBERS OF THE BOVINE AND AU-1 GENOGROUPS

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ARCHIVES OF VIROLOGY 1992年第1-4期126卷 337-342页

作者： NAKAGOMI, O KAGA, E GERNA, G SARASINI, A NAKAGOMI, T AKITA UNIV SCH MEDDEPT OCCUPAT MEDAKITA 010JAPAN UNIV PAVIA INST INFECT DISVIRUS LABI-27100 PAVIAITALY

Two human rotavirus strains, PCP 5 and MZ 58, which possessed an unusual combination of subgroup (I), serotype (3) and RNA pattern (long) were examined by RNA-RNA hybridization to determine their genogroup. While these two strains did not belong to either the Wa or the DS-1 genogroup, PCP 5 and MZ 58 possessed seven gene segments that formed hybrids with bovine rotavirus strain NCDV and four gene segments that formed hybrids with human rotavirus strain AU-1. These results suggest that PCP 5 and MZ 58 were intergenogroup reassortants formed in nature between a member of the bovine rotavirus genogroup and a member of the AU-1 genogroup.

关键词：基因病毒/遗传学核酸杂交 RNA探针/遗传学 RNA 病毒/遗传学重组遗传/遗传学轮状病毒属/遗传学动物牛

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Prevalence of, and antigenic variation in, serotype G10 rotaviruses and detection of serotype G3 strains in diarrheic calves: Implications for the origin of G10P11 or P11 type reassortant asymptomatic strains in newborn children in India

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ARCHIVES OF VIROLOGY 2002年第1期147卷 143-165页

作者： Varshney, B Jagannath, MR Vethanayagam, RR Kodhandharaman, S Jagannath, HV Gowda, K Singh, DK Rao, CD Indian Inst Sci Dept Microbiol & Cell Biol Bangalore 560012 Karnataka India Natl Dairy Dev Board Div Biotechnol Anand Gujarat India

Previous studies have shown predominant association of G10P11 type bovine rotavirus-derived reassortant strains with asymptomatic infections in newborn children in India. To understand the epidemiological and genetic basis for the origin of these strains in humans, the relative frequencies of different serotypes among bovine rotaviruses (BRVs) isolated from southern, western and central regions of the country were determined by subgroup and serotype analysis as well as nucleotide (nt) sequence analysis of the genes encoding the outer capsid proteins VP4 and VP7. Since the human G10P11 asymptomatic neonatal strain I321 possessed NSP1 from a human rotavirus, to determine its genetic origin in the bovine strains, comparative analysis of partial gene sequences from representative G10P11 strains was also carried out. The following observations were of great epidemiological significance, (i) G10P11 strains predominated in all the three regions with frequencies ranging between 55.6% and 85.2%. In contrast to the high prevalence of G6 strains in other countries, only one G6 strain was detected in this study and G8 strains represented 5.8% of the isolates, (ii) among the G10 strains, in serotyping ELISA, four patterns of reactivity were observed that appeared to correlate with the differences in electropherotypic patterns and amino acid (aa) sequence of the VP7, (iii) surprisingly, strains belonging to serotype G3 were detected more frequently (10.7%) than those of serotypes G6 and G8 combined, while strains representing the new serotype (G15) were observed in a single farm in Bangalore, and (iv) about 3.9% of the isolates were nontypeable as they exhibited high cross-reactivity to the serotyping MAbs used in the study. Comparative analysis of the VP7 gene sequence from the prototype G3 MAb-reactive bovine strain J63 revealed greatest sequence relatedness (87.6% nt and 96.0% aa) with that of serotype G3 rhesus-monkey strain RRV. It also exhibited high sequence homology with

关键词：氨基酸序列抗原变异抗原病毒碱基序列病毒壳体/遗传学病毒壳体/免疫学衣壳蛋白质类牛疾病/流行病学牛疾病/病毒学电泳聚丙烯酰氨凝胶印度/流行病学分子序列数据患病率再装配病毒/遗传学轮状病毒属/遗传学轮状病毒属/免疫学轮状病毒感染/流行病学轮状病毒感染/兽医学轮状病毒感染/病毒学序列比对序列分析 DNA 血清分型病毒非结构蛋白质类/遗传学病毒非结构蛋白质类/免疫学动物猫牛狗人类婴儿新生小鼠

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Characterization of a monoclonal antibody directed to the surface of MA104 cells that blocks the infectivity of rotaviruses

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VIROLOGY 2000年第1期273卷 160-168页

作者： López, S Espinosa, R Isa, P Merchant, MT Zárate, S Méndez, E Arias, CF Univ Nacl Autonoma Mexico Inst Biotechnol Dept Genet & Fisiol Mol Cuernavaca 62250 Morelos Mexico Univ Nacl Autonoma Mexico Fac Med Dept Microbiol & Parasitol Cuernavaca 62250 Morelos Mexico

Rhesus rotavirus (RRV) binds to sialic acid residues on the surface of target cells, and treatment of these cells with neuraminidase greatly reduces virus binding with the consequent reduction of infectivity. Variants that can efficiently infect neuraminidase-treated cells have been isolated, indicating that attachment to sialic acid is not an essential step for animal rotaviruses to infect cells. To identify and characterize the neuraminidase-resistant receptor for rotaviruses, we have isolated a hybridoma that secrets a monoclonal antibody (MAb) (2D9) that specifically blocks the infectivity of wild-type (wt) RRV and of its sialic acid-independent variant nar3, in untreated as well as in neuraminidase-treated cells. The infectivity of a human rotavirus was also inhibited, although to a lesser extent. MAb 2D9 blocks the binding of the variant to MA104 cells, while not affecting the binding of wt RRV;in addition, this MAb blocked the attachment of a recombinant glutathione S-transferase (GST)-VP5 fusion protein, but did not affect the binding of GST-VP8. Altogether these results suggest that MAb 2D9 is directed to the neuraminidase-resistane receptor. This receptor seems to mediate the direct attachment of the variant to the cell, through VP5, while the receptor is used by wt RRV for a secondary interaction, after its initial binding to sialic acid, through VP8. MAb 209 interacts specifically with the cell surface by indirect immunofluorescence, immunoelectron microscopy, and FAGS. By a solid-phase immunoisolation technique, MAb 2D9 was found to react with three proteins of ca. 47 55, and 220 kDa, which might form a complex. (C) 2000 Academic Press.

关键词：抗体特异性生物素化作用细胞系表位/免疫学表位/代谢流式细胞术荧光抗体技术间接杂交瘤/免疫学恒河猴/病毒学显微镜检查免疫电子分子量突变/遗传学 N-乙酰神经氨酸/代谢神经氨酸酶/代谢 RNA结合蛋白质类/代谢受体病毒/拮抗剂和抑制剂受体病毒/免疫学受体病毒/代谢重组融合蛋白质类/拮抗剂和抑制剂重组融合蛋白质类/代谢轮状病毒属/化学轮状病毒属/药物作用轮状病毒属/遗传学轮状病毒属/代谢变异(遗传学)/遗传学病毒非结构蛋白质类/拮抗剂和抑制剂病毒非结构蛋白质类/代谢人类小鼠

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Rotavirus

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CLINICS IN LABORATORY MEDICINE 1999年第3期19卷 691-+页

作者： Steele, JCH Med Coll Georgia Clin Microbiol Lab BAS 173B Augusta GA 30912 USA

Rotavirus is the leading cause of acute nonbacterial gastroenteritis (ANBGE) in infants and young children. In temperate climates it accounts for most cases of "winter diarrhea." Generally thought to spread by person-to-person contact, rotavirus has been implicated in some food-borne outbreaks. This article reviews current knowledge of this prevalent viral pathogen.

关键词：胃肠炎/病毒学轮状病毒属/分类轮状病毒属/遗传学轮状病毒属/致病力轮状病毒感染/诊断轮状病毒感染/治疗轮状病毒感染/病毒学人类

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Characterization of human symptomatic rotavirus isolates MP409 and MP480 having 'long' RNA electropherotype and subgroup I specificity, highly related to the P6,G8 type bovine rotavirus A5, from Mysore, India

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ARCHIVES OF VIROLOGY 2000年第7期145卷 1339-1357页

作者： Jagannath, MR Vethanayagam, RR Reddy, BSY Raman, S Rao, CD Indian Inst Sci Dept Microbiol & Cell Biol Bangalore 560012 Karnataka India

In an epidemiological study of symptomatic human rotaviruses in Mysore, India during 1993 and 1994, isolates MP409 and MP480 were isolated from two children suffering from severe, acute dehydrating diarrhea. Both isolates exhibited 'long' RNA pattern and subgroup I specificity suggesting the likelihood of their animal origin. Both isolates did not react with monoclonal antibodies (MAbs) specific for serotypes G1 to G6 as well as CIO. To determine the genetic origin of these isolates, complete nucleotide sequences of genes encoding the outer capsid proteins VP4 and VP7, nonstructural proteins NSP1 and NSP3 and viral enterotoxin protein NSP4 from MP409 and partial sequences of genes from MP480 were determined. Comparison of the 5' and 3' terminal sequences of 250 nucleotides revealed complete identity of the gene sequences in both strains suggesting that MP409 and MP480 are two different isolates of a single strain. Comparison of the nucleotide and deduced amino acid sequences of VP4, VP7, NSP1 and NSP3 of MP409 with published sequences of strains belonging to different serotypes revealed that both outer capsid proteins VP4 and VP7 and NSP1 are highly related to the respective proteins from the P6[1], G8 type bovine rotavirus A5 isolated from a calf with diarrhoea in Thailand and that the NSP3 is highly homologous to that of bovine rotaviruses. The NSP 1 protein showed greatest sequence identity with NSP4s belonging to the KUN genetic group to which NSP4s from human G2 type strains and bovine rotaviruses belong. MP409 and MP480 likely signify interspecies transmission of P6[1], G8 type strains from cattle to humans and represent the first P6[1] type rotaviruses isolated in humans. These and our previous studies on the asymptomatic neonatal strain I321 are of evolutionary and epidemiological significance in the context of close association of majority of the Indian population with cattle.

关键词：氨基酸序列基因组病毒印度分子序列数据 RNA 病毒/遗传学轮状病毒属/分类轮状病毒属/遗传学轮状病毒属/分离和提纯序列比对序列分析病毒蛋白质类/遗传学动物牛人类

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First detection of bovine group B rotavirus in Japan and sequence of its VP7 gene

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ARCHIVES OF VIROLOGY 1999年第4期144卷 805-815页

作者： Tsunemitsu, H Morita, D Takaku, H Nishimori, T Imai, K Saif, LJ Natl Inst Anim Hlth Shichinohe Res Unit Shichinohe Aomori 0392586 Japan Natl Inst Anim Hlth Hokkaido Res Stn Sapporo Hokkaido Japan Nemuro Livestock Hyg Ctr Betsukai Hokkaido Japan Ohio State Univ Ohio Agr Res & Dev Ctr Food Anim Hlth Res Program Wooster OH 44691 USA

An epizootic outbreak of diarrhea occulted in adult cows on a dairy farm in Hokkaido, Japan. One colostrum-fed calf inoculated with pooled feces of the 5 affected cows, developed mild diarrhea, and shed rotavirus-like particles which reacted with antiserum to group B rotavirus in immune electron microscopy. Cell culture immunofluorescence tests, RNA-polyacrylamide gel electrophoresis and RT-PCR confirmed that this virus was bovine group B rotavirus, which was designated the Nemuro strain. Additional 2 colostrum-deprived calves inoculated with feces of the first calf also developed diarrhea and shed virus, suggesting that this group B rotavirus might be the etiological agent of the outbreak of adult cow diarrhea. The identities of the nucleotide (nt) and deduced amino acid (aa) sequences of the Nemuro VP7 gene were high (93-95% in nt and 96-97% in aa) and low (61-63% in nt and 49-61% in aa) compared to those of the published corresponding genes from 3 bovine and 2 other mammalian (human and rat) strains of group B rotaviruses, respectively. To our knowledge, this is the first report on the presence of bovine group B rotavirus in Japan.

关键词：氨基酸序列抗原病毒/遗传学病毒壳体/遗传学衣壳蛋白质类牛疾病/流行病学牛疾病/传播牛疾病/病毒学细胞系初乳腹泻/流行病学腹泻/兽医学腹泻/病毒学疾病暴发流行/兽医学粪便/病毒学日本/流行病学分子序列数据轮状病毒属/分类轮状病毒属/遗传学轮状病毒属/分离和提纯轮状病毒感染/流行病学轮状病毒感染/传播轮状病毒感染/兽医学序列比对序列同源性氨基酸病毒脱落动物牛女(雌)性人类大鼠

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Comparative sequence analysis identified mutations outside the NSP4 cytotoxic domain of tissue culture-adapted ATCC-Wa strain of human rotavirus and a novel inter-species variable domain in its C-terminus

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ARCHIVES OF VIROLOGY 2000年第9期145卷 1789-1799页

作者： Mohan, KVK Atreya, CD US FDA Ctr Biol Evaluat & Res Sect Viral Pathogenesis & Adverse React Lab Pediat & Resp Viral DisDiv Viral Prod Bethesda MD 20892 USA

Complete nucleotide sequence of the tissue culture-adapted ATCC*-Wa strain of human rotavirus NSP4 was determined. Sequence analysis detected two alternate forms of the gene with a nucleotide difference at position 331 (A or G) in the coding sequence (NSP4-A or NSP4-G) leading to a change from neutral glutamine(97) in NSP4-A to a positively charged arginine(97) in NSP4-G originating from the same ATCC-Wa preparation. In addition to this, both forms of ATCC-Wa NSP4 revealed three mutations at nucleotide positions 88 (T to C), 142 (C to T) and 572 (G to A), when compared to the previously reported NSP4 sequence from virulent Wa strain. The former two mutations were in the coding sequence and resulted in a leucine(16) to serine(16) and a proline(34) to leucine(34) change, while the third mutation was in the 3' non-coding region of the gene. The two amino acid changes may contribute to the 'tissue culture-adaptation' of ATCC-Wa strain. The ATCC-Wa NSP4 sequence was found to differ from the previously reported NSP4 sequence of attenuated Wa strain by lacking a mutation at 133 (T to C), though the mutations at 88 and 142 were present in both strains. Furthermore, comparison of deduced amino acid sequence of NSP4 from human, bovine, porcine and simian rotavirus strains identified a seven-residue (positions 135-141) inter-species variable domain in its C-terminal region.

关键词：氨基酸序列氨基酸取代碱基序列细胞系克隆分子指导DNA的RNA聚合酶类基因组病毒等电点亮氨酸猕猴属分子序列数据突变脯氨酸轮状病毒属/化学轮状病毒属/遗传学丝氨酸猪病毒非结构蛋白质类/化学病毒非结构蛋白质类/遗传学动物牛人类

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EXCLUSIVE ASYMPTOMATIC NEONATAL INFECTIONS BY HUMAN ROTAVIRUS STRAINS HAVING SUBGROUP-I SPECIFICITY AND LONG RNA ELECTROPHEROTYPE

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ARCHIVES OF VIROLOGY 1992年第1-4期126卷 239-251页

作者： SUKUMARAN, M GOWDA, K MAIYA, PP SRINIVAS, TP KUMAR, MS AIJAZ, S REDDY, RR PADILLA, L GREENBERG, HB RAO, CD INDIAN INST SCI DEPT MICROBIOL & CELL BIOL BANGALORE 560012 KARNATAKA INDIA MSR MED COLL HOSP BANGALORE INDIA ST MARTHAS HOSP BANGALORE INDIA STANFORD UNIV MED CTR SCH MED DIV GASTROENTEROL STANFORD CA 94305 USA INDIAN INST SCI CTR GENET ENGN BANGALORE 560012 KARNATAKA INDIA

A large number of stool specimes, of healthy new born infants, collected from various hospitals and clinics in Bangalore City, India, have been examined for the presence of asymptomatic rotaviral excretion. Out of 370 samples analysed during a three year period from 1988 to 1991, 133 specimens (36%) were positive for rotavirus RNA. All these asymptomatic neonatal strains, without exception, showed "long" RNA pattern, but subgroup I specificity. Serotype analysis by ELISA or by hybridization with serotype-specific probes indicated that these strains probably represent a new serotype in newborn children. We f-ind an exclusive association of human rotaviruses having "long" RNA pattern and subgroup I specificity with asymptomatic neonatal infections in contrast to the earlier observations of association of such unusual strains with acute gastroenteritis in young children.

关键词：抗原病毒病毒壳体/遗传学病毒壳体/免疫学衣壳蛋白质类 DNA探针/遗传学酶联免疫吸附测定粪便/微生物学印度/流行病学 RNA 病毒/遗传学轮状病毒属/分类轮状病毒属/遗传学轮状病毒属/免疫学轮状病毒属/分离和提纯轮状病毒感染/流行病学轮状病毒感染/微生物学血清分型人类婴儿新生

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CODING ASSIGNMENTS OF THE GENOME OF ADULT DIARRHEA ROTAVIRUS

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ARCHIVES OF VIROLOGY 1992年第1-4期125卷 53-69页

作者： FANG, ZY MONROE, SS DONG, H PENARANDA, M WEN, L GOUVEA, V ALLEN, JR HUNG, T GLASS, RI CHINESE ACAD PREVENT MED INST VIROL BEIJING PEOPLES R CHINA

Adult diarrhea rotavirus (ADRV) has caused epidemics of diarrhea in China since 1982 and remains the only group B rotavirus associated with widespread disease in humans. We recently characterized the proteins of ADRV and have now proceeded to identify the gene segments encoding each protein. Viral RNA transcripts were synthesized in vitro with the endogenous viral RNA polymerase and separated by electrophoresis in agarose. The individual transcripts were translated in a cell-free system using nuclease-treated rabbit reticulocyte lysates. The translation products were compared with polypeptides found in purified virus and were characterized by SDS-PAGE, immunoprecipitation, and Western blot analysis using antisera to double- and single-shelled virions, virus cores, and monoclonal antibodies. Furthermore, individual RNA transcripts were hybridized to total dsRNA to determine their genomic origin. Based on this analysis, the core polypeptides VP1, VP2 and VP3 are encoded by segments 1, 2, and 3, respectively. The main polypeptides in the inner capsid, VP6, and the outer capsid, VP4 and VP7, are encoded by segments 6, 4, and 8 respectively. Segments 5, 7, and 9 code for 60, 45, and 30 kDa nonstructural polypeptides. Two other nonstructural polypeptides (24 and 25 kDa) are derived from gene segment 11. Gene segment 10 codes for a 26 kDa polypeptide. that is precipitated with serum to ADRV and may be a structural protein VP9. With this exception, gene coding assignments of ADRV are comparable to those of the group A rotaviruses. Our results have clear implications for further work in cloning, sequencing, and expression genes of ADRV and can provide direction towards understanding the origin and the evolution of this virus.

关键词：无细胞系统染色体图腹泻/微生物学遗传密码基因组病毒糖蛋白类/遗传学肽生物合成肽类/遗传学蛋白质生物合成 RNA 信使/遗传学 RNA 病毒/遗传学轮状病毒属/遗传学病毒蛋白质类/生物合成病毒蛋白质类/遗传学病毒粒子/遗传学动物人类兔

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