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Characterisation of New Zealand isolates of infectious bursal disease virus

ARCHIVES OF VIROLOGY 2001年第8期146卷 1571-1580页

作者： Chai, YF Christensen, NH Wilks, CR Meers, J Massey Univ Inst Vet Anim & Biomed Sci Palmerston North New Zealand Pacific Vet Ltd Christchurch New Zealand

Isolates of infectious bursal disease virus (IBDV) were obtained from domestic poultry in New Zealand in 1997 and 1998. An in-vivo pathogenicity study carried out in specific pathogen free (SPF) chickens demonstrated the low virulence of one of the virus isolates. The nucleotide sequences of the hypervariable region of the VP2 gene of two isolates were determined and compared with published sequences of strains from other countries. The deduced amino acid sequence of the two New Zealand IBDV isolates showed 100% identity with each other, suggesting that little genetic drift had occurred. Phylogenetic analysis showed that the New Zealand isolates were more closely related to two attenuated IBDV strains (Cu1 and PBG98) than to classical (STC and 52/70), very virulent (DV86), variant (variant E) or Australian (002-73) strains. The results support the hypothesis that an attenuated strain of the virus was inadvertently introduced into the NZ poultry population in 1993.

关键词：氨基酸序列双RNA病毒科感染/兽医学双RNA病毒科感染/病毒学鸡 DNA 互补传染性腔上囊病病毒/遗传学传染性腔上囊病病毒/分离和提纯传染性腔上囊病病毒/致病力分子序列数据新西兰家禽疾病/病毒学 RNA 病毒/分离和提纯逆转录聚合酶链反应序列分析 DNA 病毒结构蛋白质类/化学病毒结构蛋白质类/遗传学毒力动物

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Molecular epidemiology of TT virus (TTV) and characterization of two novel TTV genotypes in Indonesia

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ARCHIVES OF VIROLOGY 2001年第7期146卷 1249-1266页

作者： Muljono, DH Nishizawa, T Tsuda, F Takahashi, M Okamoto, H Jichi Med Sch Div Immunol Minami Kawachi Tochigi 3290498 Japan Jichi Med Sch Div Mol Virol Minami Kawachi Tochigi Japan Eijkman Inst Mol Biol Infect Dis Lab Jakarta Indonesia Toshiba Gen Hosp Dept Med Sci Tokyo Japan

The prevalence of TT virus (TTV) DNA among 244 healthy individuals in 23 cities on 12 islands in Indonesia was determined by polymerase chain reaction (PCR) with primers derived from the coding region (N22), which can detect TTV DNA of genotypes 1-6. By N22 PCR, TTV DNA was detected in 102 (42%) individuals. The amplified PCR products were molecularly cloned and three clones each were subjected to sequence analysis. Three hundred one (98%) of the 306 TTV clones were classified into genotype 1, 2 or 3, and none into genotypes 4-6. The remaining five clones from two individuals (Kt-08 and Kt-10) on Kutai, Kalimantan Island, differed by > 30% from known TTV isolates of all 21 genotypes and were tentatively classified into genotypes 22 and 23, respectively. Using primers specific for the new TTV genotype 22 or 23, TTV genotype 22 was detected significantly more frequently in Kutai than in the other 22 cities (41% vs. 5%, P < 0.001). TTV genotype 23 was restricted to Kutai (17% vs. 0%, P < 0.001), suggesting the indigenous nature of this genotype. Analysis of two TTV isolates (Kt-08F and Kt-10F) demonstrated the extreme diversity of TTV and the preservation of the genomic organization and transcription profile.

关键词：克隆分子 DNA病毒感染/流行病学 DNA病毒感染/病毒学 DNA 病毒/遗传学 DNA 病毒/分离和提纯进化分子基因型印度尼西亚/流行病学分子流行病学分子序列数据核酸类/分离和提纯开放读码框架种系发生聚合酶链反应/方法序列同源性氨基酸序列同源性核酸输血传播病毒/分类输血传播病毒/遗传学输血传播病毒/分离和提纯成年人女(雌)性人类男(雄)性中年人

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Promoter studies of a polydnavirus gene from Cotesia rubecula (Hym: Braconidae)

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ARCHIVES OF VIROLOGY 2001年第10期146卷 1979-1989页

作者： Asgari, S Schmidt, O Univ Adelaide Dept Appl & Mol Ecol Glen Osmond SA 5064 Australia

The Cotesia rubecula polydnavirus gene, CrV1, is expressed in a highly transient fashion. Within four hours after egg deposition and virus infection, tissues of the host caterpillar, Pieris rapae, express high levels of the transcript. Twelve hours after infection no transcripts are visible. We have previously shown that the CrV1 secreted protein is mainly produced in host haemocytes. In haemocytes, immune functions such as phagocytosis and cell spreading are abolished by destabilization of the cell cytoskeleton. To test whether the observed down-regulation of CrV1 transcripts is mediated by transcriptional control or by other factors, such as the disruption of cytoskeleton in CrV1-inactivated cells, we cloned the promoter and the 3' untranslated region of the CrV1 gene to study CrV1 expression. The promoter region of the CrV1 gene was cloned into baculovirus expression systems along with the CAT reporter gene. Molecular analyses showed that the CAT gene under the control of CrV1 promoter is expressed as early as 2 h post infection and continues until late phase of infection suggesting that down-regulation of CrV1 expression in host haemocytes is perhaps mediated by post-transcriptional mechanisms.

关键词：碱基序列细胞培养的氯霉素O-乙酰转移酶/代谢 DNA 病毒/化学 DNA 病毒/分离和提纯基因报告分子序列数据多DNA病毒科/遗传学启动区遗传斜纹夜蛾属黄蜂/病毒学动物

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Partial characterisation of citrus leaf blotch virus, a new virus from Nagami kumquat

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ARCHIVES OF VIROLOGY 2001年第2期146卷 357-368页

作者： Galipienso, L Vives, MC Moreno, P Milne, RG Navarro, L Guerri, J Inst Valenciano Invest Agr E-46113 Moncada Valencia Spain CNR Ist Fitovirol Applicata I-10126 Turin Italy

Citrus leaf blotch virus (CLBV) was purified from leaves of Nagami kumquat SRA-153 that showed bud union crease when propagated on Troyer citrange. Virions were filamentous particles (960 x 14 nm) containing a 42 kDa protein and a single-stranded RNA (ssRNA) of about 9,000 nt (Mr 3 x 10(6)). Infected tissue contained three species of double-stranded RNA (dsRNA) of Mr 6, 4.5 and 3.4 x 10(6). The nucleotide sequence of several complementary DNA (cDNA) clones showed significant similarities with replication-related proteins from plant filamentous viruses in several genera. A digoxigenin-labelled probe from one of these cDNA clones hybridised in Northern blots with ssRNA from virions and with the three dsRNA species, suggesting that the ssRNA is the genomic RNA of the virus, the largest dsRNA is its replicative form, and the two smaller dsRNAs probably replicative forms of 5' co-terminal subgenomic RNAs. CLBV was also detected in several citrus cultivars from Spain and Japan including Navelina sweet orange field trees propagated on Troyer citrange showing bud union crease;however, no virus could be detected in other citrus trees with similar symptoms. This indicates that CLBV is not restricted to kumquat SRA-153, but its involvement in causing the bud union disorder remains unclear.

关键词： RNA 双链/遗传学 RNA 双链/分离和提纯 RNA 病毒/遗传学 RNA 病毒/分离和提纯病毒粒子/遗传学

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Diagnosis of foot-and-mouth disease by RT-PCR: use of phylogenetic data to evaluate primers for the typing of viral RNA in clinical samples

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ARCHIVES OF VIROLOGY 2001年第12期146卷 2421-2434页

作者： Reid, SM Ferris, NP Hutchings, GH De Clercq, K Newman, BJ Knowles, NJ Samuel, AR AFRC Inst Anim Hlth Pirbright Lab Vesicular Dis Grp Woking GU24 0NF Surrey England Vet & Agrochem Res Ctr Brussels Belgium

The results of type-specific RT-PCR diagnostic assays on foot-and-mouth disease (FMD) viruses in clinical samples were mapped onto serotype-specific dendrograms representing the degree of nucleotide sequence variation between the FMD virus isolates. This novel approach assisted the selection of suitable PCR primer sets for the diagnosis of FMD virus isolates belonging to different topotypes within each serotype. These interpretations were qualified by using a universal (FMD virus group) specific primer to confirm that FMD virus RNA had been extracted from the samples under investigation. The analyses showed that the design of primer sets for the detection of FMD virus serotypes O, A, Asia 1, SAT 1 and SAT 3 were generally satisfactory, as most virus isolates within the major virus sub-groupings were successfully detected. However, the FMD virus serotype C and SAT 2 specific primers were less efficient as certain virus sub-groups were not detected. This identified the need for additional or alternative primers to improve RT-PCR procedures for more comprehensive detection of divergent virus strains within these serotypes. There were some examples where not all virus isolates from the same outbreak reacted with particular type-specific primers which suggested that either further minor refinements may be necessary in the primer design or that there were shortcomings in the RT-PCR methodology.

关键词： DNA引物口蹄疫/诊断口蹄疫/病毒学口蹄疫病毒/分类口蹄疫病毒/遗传学口蹄疫病毒/分离和提纯种系发生 RNA 病毒/分析 RNA 病毒/分离和提纯逆转录聚合酶链反应敏感性与特异性序列分析 DNA 血清分型动物

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Detection and typing of human papillomavirus DNA in penile carcinoma - Evidence for multiple independent pathways of penile carcinogenesis

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AMERICAN JOURNAL OF PATHOLOGY 2001年第4期159卷 1211-1218页

作者： Rubin, MA Kleter, B Zhou, M Ayala, G Cubilla, AL Quint, WGV Pirog, EC Cornell Univ Weill Med Coll Dept Pathol New York NY 10021 USA Univ Michigan Dept Pathol Ann Arbor MI 48109 USA Delft Diagnost Lab Delft Netherlands Baylor Coll Med Dept Pathol Houston TX 77030 USA Univ Nacl Asuncion Dept Pathol Asuncion Paraguay Univ Amsterdam Acad Med Ctr NL-1105 AZ Amsterdam Netherlands

To clarify the role of human papillomavirus (HPV) in penile cancer we evaluated the prevalence of HPV DNA in different histological subtypes of penile carcinoma, dysplasia, and condyloma using a novel, sensitive SPF10 HPV polymerase chain reaction assay and a novel genotyping line probe assay, allowing simultaneous identification of 25 different HPV types. Formalin-fixed, paraffin-embedded tissue samples were collected from the United States and Paraguay. HPV DNA was detected in 42% cases of penile carcinoma, 90% cases of dysplasia, and 100% cases of condyloma. There were significant differences in HPV prevalence in different histological cancer subtypes. Although keratinizing squamous cell carcinoma and verrucous carcinoma were positive for HPV DNA in only 34.9 and 33.3% of cases, respectively, HPV DNA was detected in 80% of basaloid and 100% of warty tumor subtypes. There was no significant difference in BPV prevalence between cases from Paraguay and the United States. In conclusion, the overall prevalence of BPV DNA in penile carcinoma (42%) is lower than that in cervical carcinoma (similar to 100%) and similar to vulvar carcinoma (similar to 50%). In addition, specific histological subtypes of penile cancer-basaloid and warty - are consistently associated with HPV, however, only a subset of keratinizing and verrucous penile carcinomas is positive for BTV DNA, and thus these two tumor groups seem to develop along different pathogenetic pathways.

关键词：癌鳞状细胞/病毒学尖锐湿疣/病毒学 DNA 病毒/分类 DNA 病毒/分离和提纯乳头状瘤病毒科/分类乳头状瘤病毒科/遗传学乳头状瘤病毒感染/病理学乳头状瘤病毒感染/病毒学阴茎肿瘤/病毒学阴茎/病理学阴茎/病毒学肿瘤病毒感染/病理学肿瘤病毒感染/病毒学成年人老年人老年人 80以上人类男(雄)性中年人

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Detection of TT virus DNA in liver biopsies by in situ hybridization

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AMERICAN JOURNAL OF PATHOLOGY 2000年第4期156卷 1227-1234页

作者： Rodríguez-Iñigo, E Casqueiro, M Bartolomé, J Ortiz-Movilla, N López-Alcorocho, JM Herrero, M Manzarbeitia, F Oliva, H Carreño, V Fdn Jimenez Diaz Dept Hepatol Madrid 28040 Spain Fdn Jimenez Diaz Dept Pathol Madrid 28040 Spain Fdn Estudio Hepatitis Virales Madrid Spain

A novel hepatitis-associated virus named TT virus (TTV) has been isolated. However, its hepatotropism has not been proven. We have retrospectively analyzed the presence of TTV-DNA by polymerase chain reaction (PCR) and in situ hybridization in liver biopsies from 30 patients with liver disease (15 TTV-DNA-positive and 15 TTV-DNA-negative in serum), and prospectively in serum and liver from eight patients with normal liver histology. TTV-DNA was detected by PCR in the liver from the 15 patients with serum TTV-DNA and in serum and liver of two of the eight patients without liver disease. TTV-DNA titers in liver were 10 times higher than in. serum, although no correlation between TTV-DNA. titers in serum and liver mere observed. In situ hybridization shows positive signals in the hepatocytes of the 17 patients infected by TTV but in none of the TTV-DNA-negative patients by PCR. No morphological changes were observed in the hepatocytes showing hybridization signals. The percentage of positive hepatocytes ranged from 2.1% to 30% and correlated with the TTV-DNA titers in liver (r = 0.54;P = 0.037). In conclusion, our results show that TTV is able to infect liver cells although they do not support a role for TTV in causing liver disease.

关键词：碱基序列/遗传学活组织检查圆环病毒科/遗传学 DNA病毒/遗传学 DNA 病毒/分离和提纯原位杂交肝/病理学肝/病毒学肝疾病/病理学肝疾病/病毒学成年人女(雌)性人类男(雄)性中年人

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Comparison of nucleotide sequences between northern and southern Philippine isolates of rice grassy stunt virus indicates occurrence of natural genetic reassortment

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VIROLOGY 2000年第1期266卷 26-32页

作者： Miranda, GJ Azzam, O Shirako, Y Univ Tokyo ANESC Asian Ctr Bioresources & Environm Sci Bunkyo Ku Tokyo 1138657 Japan Univ Tokyo Grad Sch Agr Life Sci Bunkyo Ku Tokyo 1138657 Japan Int Rice Res Inst Makati 1271 Philippines

Rice grassy stunt virus is a member of the genus Tenuivirus, is persistently transmitted by a brown planthopper, and has occurred in rice plants in South, Southeast, and East Asia (similar to North and South America). We determined the complete nucleotide (nt) sequences of RNAs 1 (9760 nt), 2 (4069 nt), 3 (3127 nt), 4 (2909 nt), 5 (2704 nt), and 6 (2590 nt) of a southern Philippine isolate from South Cotabato and compared them with those of a northern Philippine isolate from Laguna (Toriyama et al., 1997 1998). The numbers of nucleotides in the terminal untranslated regions and open reading frames were identical between the two isolates except for the 5' untranslated region of the complementary strand of RNA 4. Overall nucleotide differences between the two isolates were only 0.08% in RNA 1, 0.58% in RNA 4, and 0.26% in RNA 5, whereas they were 2.19% in RNA 2, 8.38% in RNA 3, and 3.63% in RNA 6. In the intergenic regions, the two isolates differed by 9.12% in RNA 2, 11.6% in RNA 3, and 6.86% in RNA 6 with multiple consecutive nucleotide deletion/insertions, whereas they differed by only 0.78% in RNA 4 and 0.34% in RNA 5. The nucleotide variation in the intergenic region of RNA 6 within the South Cotabato isolate was only 0.33%. These differences in accumulation of mutations among individual RNA segments indicate that there was genetic reassortment in the two geographical isolates;RNAs 1, 4, and 5 of the two isolates came from a common ancestor, whereas RNAs 2, 3, and 6 were from two different ancestors. (C) 2000 Academic Press.

关键词：氨基酸序列 DNA 互补/遗传学分子序列数据突变水稻/病毒学菲律宾植物病毒/遗传学植物病毒/分离和提纯 RNA病毒/遗传学 RNA病毒/分离和提纯 RNA 病毒/遗传学 RNA 病毒/分离和提纯再装配病毒/遗传学再装配病毒/分离和提纯序列比对序列分析 DNA 变异(遗传学) 病毒蛋白质类/化学病毒蛋白质类/遗传学

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Human herpesvirus-8-associated disseminated angiosarcoma in an HIV-seronegative woman: Report of a case and limited case-control virologic study in vascular tumors

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AMERICAN JOURNAL OF MEDICINE 2000年第8期108卷 660-664页

作者： Remick, SC Patnaik, M Ziran, NM Liegmann, KR Dong, J Dowlati, A Yao, Y Abdul-Karim, FW Giam, CZ Case Western Reserve Univ Sch Med Dept Med Cleveland OH 44106 USA Case Western Reserve Univ Sch Med Dept Pathol Cleveland OH 44106 USA Specialty Labs Santa Monica CA USA Uniformed Serv Univ Hlth Sci Dept Microbiol & Immunol Bethesda MD 20814 USA

In 1994, Chang and colleagues (1) used representational difference analysis to show that a unique DNA sequence was associated with Kaposi□s sarcoma in patients with the acquired immunodeficiency syndrome (AIDS). These sequences, which are of nonhuman origin, are closely homologous to minor capsid and tegument protein genes of the Gammaherpesvirinae (rhadinovirus subgroup), herpesvirus samirii, and Epstein-Barr virus. This observation was subsequently extended to patients with Kaposi□s sarcoma who did not have human immunodeficiency virus (HIV) infection (2). Since its initial discovery, the Kaposi□s sarcoma–associated herpesvirus/human herpesvirus-8 (KSHV/HHV-8) virus has been linked to two other lymphoproliferative disorders: multicentric Castleman□s disease (3,4) and a newly recognized disease, body-cavity–based lymphoma or primary effusion lymphoma (5–7). Human herpesvirus-8 infection of bone marrow dendritic cells in HIV-seronegative patients with multiple myeloma has also been reported (8,9), although the initial observations have not been replicated (10). Current epidemiologic evidence indicates that human herpesvirus-8 infection has little or no effect on common cancer types or sites, including multiple myeloma (11–13). A few reports, however, have identified the virus in diseases associated with vascular hyperplasia, including follicular B-cell lymphoma (14), benign and angioimmunoblastic lymphadenopathy (15), angiolymphoid hyperplasia with eosinophilia (16), and angiosarcoma (17).

关键词：乳腺肿瘤/病毒学病例对照研究 DNA 病毒/分离和提纯致命性结局荧光抗体技术巨淋巴结增生/病毒学 HIV血清阴性血管肉瘤/病毒学疱疹病毒科感染/并发症疱疹病毒8型人/遗传学疱疹病毒8型人/分离和提纯肝肿瘤/病毒学肉瘤卡波西/病毒学肿瘤病毒感染/并发症女(雌)性人类中年人

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Enhanced replication contributes to enrichment of hepatitis B virus with a deletion in the core gene

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VIROLOGY 2000年第2期273卷 286-299页

作者： Günther, S Piwon, N Jung, A Iwanska, A Schmitz, H Will, H Bernhard Nocht Inst Trop Med D-20359 Hamburg Germany Univ Hamburg Heinrich Pette Inst Expt Virol & Immunol D-20251 Hamburg Germany

Accumulation in immunosuppressed patients of hepatitis a virus (HBV) with a deletion in the C gene is associated with severe liver disease. The aim of this study was to determine the phenotype of such genomes in vitro. Four C gene fragments with different types of deletions were inserted in the context of a wild-type genome and tested by transfection into HuH7 cells. The deletions did not influence mRNA and surface protein levels. Truncated C gene translation products were expressed only from variants with in-frame deletions, whereas full-length polymerase was expressed from all variants at a similar or higher level than in wild-type virus. None of the variants was competent for autonomous replication;however, they produced 2- to 4.5-fold more progeny DNA than wild-type HBV when sufficiently complemented with wild-type core protein. Similarly, when variant and wild-type DNA were cotransfected in different ratios, the variants produced 2- to 5-fold more progeny DNA relative to the wild-type;this enrichment required the expression of the viral polymerase in c/s. The mechanism of enrichment depended on the percentage of variant in the transfected DNA mixture. When the transfected DNA contained a small percentage of variant, enhanced replication of the variant accompanied by no or little suppression of wild-type replication was seen. Accordingly, overall production of progeny virus was slightly increased. At a high percentage of variant DNA, replication of both variant and wild-type decreased, probably due to a shortage of wild-type core protein. In conclusion, emergence of C gene deletion variants in vivo may be due to enhanced replication mediated at the level of encapsidation or reverse transcription. If the variants constitute a small part of the ccc DNA, they can be fully trans-complemented by wild-type virus which may increase the overall virus production. (C) 2000 Academic Press.

关键词：细胞培养的克隆分子 DNA复制 DNA 病毒/生物合成 DNA 病毒/化学 DNA 病毒/分离和提纯基因组病毒肝炎表面抗原乙型/生物合成肝炎e抗原乙型/生物合成肝炎病毒乙型/遗传学肝炎病毒乙型/生理学表型多态性限制性片段长度 RNA 信使/代谢 RNA 病毒/代谢转染病毒复制人类

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