Electronic Library of Scientific Literature Electronic Library of Scientific Literature
Volume 43 / December 1999 / number 6
S. SCHüNEMANN, C. MAINKA, M.H. WOLFF
Institute of Microbiology and Virology, University of Witten/Herdecke, Stockumer Str. 10, D-58448 Witten, Germany
Summary. - Patients suffering from postherpetic neuralgia (PHN) were investigated whether varicella-zoster virus (VZV) DNA or RNA could be detected in their peripheral blood mononuclear cells (PBMCs). Altogether 16 samples were tested by standard polymerase chain reaction (PCR) for open reading frame (ORF) 14 and ORF 29, standard and nested PCR for ORF 63, and isothermal transcription-based nucleic acid amplification (NASBA) for ORF 63 and ORF 68. By these methods neither VZV DNA nor VZV RNA could be detected. The obtained results are in contrast to those of other authors (Vafai et al., 1988; Mahalingam et al., 1995) but support the hypothesis of Bennett (1994) and Kost and Straus (1996) proposing that PHN is not caused by acute VZV replication but a consequence of neuronal damage accompanying replication of VZV in ganglia during zoster episodes.
Key words: varicella-zoster virus; postherpetic neuralgia; PCR; NASBA
Acta virologica 43: 337 - 340, 1999
A. Akin, T.L. Lin, C.C. Wu
Department of Veterinary Pathobiology, School of Veterinary Medicine, Purdue University, 1175 ADDL, West Lafayette, IN 47907-1175, USA
Summary. - Five hammerhead-type ribozymes were designed and cloned to cleave infectious bursal disease virus (IBDV) RNA and inhibit protein synthesis from cloned full-length viral cDNA genes. Two ribozymes (R1 and R2) were directed to the large viral RNA segment gene and three ribozymes (R3, R4, and R5) were directed to the small viral RNA segment gene. Targets for the ribozymes were produced from cloned full-length coding regions of both small and large viral RNA segment genes. Ribozymes and their corresponding targets were synthesized as in vitro transcripts. Despite several attempts at different temperatures, no cleavage of viral RNA transcripts with four of the ribozymes (R1, R2, R3, and R5) was observed. One of the ribozymes, R4, was effective in cleaving the viral RNA polymerase gene transcripts in a magnesium-dependent manner. Ribozyme R4 caused 82% reduction in the synthesis of the viral RNA polymerase gene product. The inhibition was specific since there was no change in the rate of synthesis of the Escherichia coli beta-galactosidase protein. The results suggest that ribozyme R4 can be used as potential anti-IBDV agent. It was also demonstrated that the hammerhead-type ribozymes can cleave sites other than conventional GUC sequence motif as the cleavage site of ribozyme R4 had GUU motif which conformed to the NUX consensus motif.
Key words: infectious bursal disease virus; double-stranded RNA; ribozyme;
gene therapy
Acta virologica 43: 341 - 347, 1999
J. Fránová-Honetšlegrová, I. Mráz, J. Nebesářová, M. Šíp
Department of Plant Virology, Institute of Plant Molecular Biology, Academy of Sciences
of the Czech Republic, Branišovská 31, 370 05 České Budějovice;
Laboratory of Electron Microscopy, Institute of Parasitology, Academy of Sciences of the
Czech Republic, České Budějovice;
Faculty of Biology, University of South Bohemia, České Budějovice; Czech Republic
Summary. - Leaves of Fragaria ananassa Duch. cv. Redgauntlet with mottle and mild dwarf symptoms were grafted onto F. vesca indicator clones. The youngest leaves developed specific vein banding pattern located preferentially on secondary veins near the edge of the leaves. Electron microscopy of ultrathin sections and negatively stained purified virus preparations from symptom-bearing strawberry leaves revealed presence of different-sized isometric virions. Particles of about 50 nm and 23 nm in diameter were identified as strawberry vein banding virus (SVBV) and tobacco necrosis virus (TNV) D strain. Based on results of electron microscopy, DNA hybridization, enzyme-linked immunosorbent assay (ELISA), and DNA sequencing we propose that the anomalous “leaf edge vein banding” symptoms are caused by a mixed virus infection with SVBV and other viruses such as TNV.
Key words: Fragaria; mixed virus infection; tobacco necrosis virus;
ultrastructure; strawberry vein banding virus; caulimovirus
Acta virologica 43: 349 - 355, 1999
L.J. DA COSTA, M.A. REBELLO
Institute of Biophysics, C.C.F, Rio de Janeiro;
Departament of Virology, Institute of Microbiology, P.P.G., Federal University of Rio de
Janeiro, Rio de Janeiro CEP 21941-590, Brazil
Summary. - Brefeldin A (BFA), a fungal metabolite that blocks transport of newly synthesized proteins from the endoplasmic reticulum, was found to inhibit Mayaro virus replication. At the concentration of 0.05 µg/ml, the yield of the virus was inhibited by 94% in Aedes albopictus cells and by 99.5% in Vero cells. Treatment of A. albopictus cells with BFA did not inhibit the virus protein synthesis. However, this compound drastically reduced viral protein synthesis in Vero cells. The inhibitory effect progressively declined when BFA was added at late times post infection (p.i.). The effect of BFA on protein glycosylation is discussed.
Key words: Mayaro virus; Brefeldin A; Aedes albopictus cells; Vero cells;
protein synthesis
Acta virologica 43: 357 - 360, 1999
L. YU, Y. LI, G. GU, Y. WANG, B. ZHOU, C. AN, Z. CHEN
College of Life Science, Peking University, P.O. Box 14, Beijing 100871, P.R. of China
Summary. - Based on stability of double-stranded (ds) RNA, a new, fast, sensitive, and specific method for detection of genomic rice dwarf virus (RDV) dsRNA by molecular hybridization was developed. In contrary to the commonly used, standard Northern blot analysis, dsRNA is denatured in the immobilized state on the blot. Therefore, risk of degradation of single-stranded (ss) RNA by ribonuclease (RNase) during sample preparation, electrophoresis and blotting is eliminated. This method overcomes disadvantage of incomplete denaturation of dsRNA in Northern blot analysis. In conclusion, the newly developed method is reliable, sensitive, very specific, and gives a low background. The entire procedure is also less time consuming; it can be completed within 2-3 days. The new method may be regarded as a modification of the standard Northern blot analysis.
Key words: rice dwarf virus; phytoreovirus; double-stranded RNA; detection;
hybridization
Acta virologica 43: 361 - 365, 1999
J.S. Nandi
Department of Virology, Division of Pathology, University College London Medical School, London, United Kingdom
Summary. - The unintegrated viral DNA found in human immunodeficiency virus (HIV) infection includes linear and circular forms. The circular unintegrated viral DNA (CUVD) could be of either 1-long terminal repeat (LTR) or 2-LTR form. Inverse primers from nef (upstream) and gag (downstream) gene sequences of HIV-1 genome were designed to span the LTR circle junction. CUVD was assayed in unstimulated, quiescent persistently infected cell lines 8E5, HIIIB, and GB8, as well as in peripheral blood lymphocytes (PBLs) of HIV-1- infected patients by nested PCR in a cross sectional study. CUVD in the infected cell lines (in vitro) was predominantly of 2-LTR form in 8E5 and GB8 cells, while in HIIIB cells, there was besides 1-LTR and 2-LTR an additional, intermediate form. In vivo, CUVD was predominantly of 1-LTR form. The possibility of using CUVD, an early phenomenon in the virus replication, as an additional postpenetration, preintegration marker of HIV infection is discussed.
Key words: human immunodeficiency virus 1; unintegrated circular DNA; cell lines;
peripheral blood lymphocytes; nested polymerase chain reaction
Acta virologica 43: 367 - 372, 1999
V. Kaden, H. Steyer, G. Strebelow, E. Lange, P. Hübert, P. Steinhagen
Bundesforschungsanstalt für Viruskrankheiten der Tiere, Institut für
Infektionsmedizin, Friedrich-Loeffler-Institute Insel Riems, Boddenblick 5a, D-17498 Insel
Riems, Germany;
Lebensmittel- und Veterinäruntersuchungsamt des Landes Schleswig-Holstein, Neumünster,
Germany
Summary. - The effectiveness of virus isolation, commercial antigen enzyme-linked immunosorbent assay (ELISA), reverse transcriptase-polymerase chain reaction (RT-PCR), and flow cytometry in detection of a low- virulent classical swine fever virus (CSFV) in blood in the early period of infection was evaluated. Domestic pigs at the age of 6-8 weeks and young wild boars were inoculated with a low-virulent field isolate of CSFV originating from a wild boar. This virus induced serious clinical reaction in only one pig which was naturally infected with Pasteurella multocida. Nine of 13 infected domestic pigs showed viremia. All infected weanling pigs were found viraemic by virus isolation on day 6 post infection (p.i.) but virus-free by RT-PCR. The flow cytometry was apparently not as sensitive as the virus isolation. Two young wild boars infected with the virus were viremic only for the first 2 days p.i. Virus isolation and RT-PCR were of similar sensitivity. Three different commercial antigen ELISAs used were not able to detect viral antigen in any animal.
Key words: classical swine fever virus; low virulence; virus detection; virus
isolation; ELISA; RT-PCR; flow cytometry
Acta virologica 43: 373 - 380, 1999
A. HUSSEIN, V. PÄTOPRSTÝ, R. TOMAN
Institute of Virology, Slovak Academy of Sciences, Dúbravská cesta 9, 842 45
Bratislava;
Institute of Chemistry, Slovak Academy of Sciences, Bratislava, Slovak Republic
Summary. - A lipopolysaccharide (LPS) of Chlamydia psittaci PK 5082 strain associated with enzootic abortion in ewes was isolated from embryonated hen eggs-grown elementary bodies (EBs) by a phenol/water procedure. Compositional analyses revealed the presence of 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo), glucosamine (GlcN), phosphorus, and fatty acids in a molar ratio of 2.6:2.0:2.4:4.4. GlcN was the only amino sugar detected. Methylation analysis of the LPS confirmed the presence of a Kdo trisaccharide proximal to lipid A having the structure Kdo-(2->8)-Kdo-(2->4)-Kdo, which appears to be a highly conserved region in native chlamydial LPSs. The complex fatty acid composition revealed the presence of ten different straight or branched (iso and anteiso) nonhydroxy fatty acids and thirteen 3-hydroxy fatty acids. The major nonhydroxy fatty acid was icosanoic acid and the most prominent 3-hydroxy fatty acid was 3-hydroxyicosanoic acid followed by 3-hydroxy-18-methylicosanoic acid. The 3-hydroxy fatty acids represented more than two thirds of the total fatty acid content and most of them were bound in amide linkages. In contrast, most nonhydroxy fatty acids were ester-linked. It appears that LPSs from various chlamydial species differ in fatty acid composition and distribution.
Key words: Chlamydia psittaci; lipopolysaccharide; chemical composition; structure;
reinvestigation
Acta virologica 43: 381 - 386, 1999
K.R.P. Sivarama, E. Sreekumar, T.J. Rasool
Indian Veterinary Research Institute, Hebbal, Bangalore 560 024, India
Summary. - Bovine herpesvirus 1 (BHV-1) has been reported from the Indian subcontinent in late 70’s. In order to identify the origin of an Indian isolate of BHV-1 (IBR/H 167 VS) and its molecular relationship to known strains of BHV-1, a 680 bp region of the glycoprotein gene gIV was amplified by polymerase chain reaction (PCR), cloned and sequenced. Comparison of this sequences with the corresponding one of an European strain of BHV-1 (Cooper) revealed more than 99% nucleotide (nt) homology. We conclude that the Indian isolate under study does not differ from the Cooper strain regarding the gIV gene.
Key words: bovine herpesvirus 1; glycoprotein gIV gene; cloning; sequencing
Acta virologica 43: 387 - 389, 1999
N. ČEŘOVSKÁ, T. MORAVEC, M. FILIGAROVÁ, H. RYŠLAVÁ, J. GROSCLAUDE
Institute of Experimental Botany, Academy of Czech Republic, Na Karlovce 1, 160 00
Prague 6;
Department of Biochemistry, Faculty of Natural Sciences, Charles University, Prague, Czech
Republic;
INRA, Virologie-Immunologie Domaine, Jouy-en-Josas, France
Summary. - Eight isolates of potato virus Y NTN strain (PVY-NTN) of different origin were studied by means of monoclonal antibodies (MAbs) in non-competitive and competitive enzyme-linked immunosorbent assay (ELISA), and by immunoblot analysis of the viral coat protein (CP). As the MAbs reacted with the denatured viral CP, their epitopes must be continuous. The ELISA data demonstrate that the epitopes are topologically different. The epitopes may be located on the N-terminal part of CP as showed its partial amino acid sequencing and the immunoblot analysis.
Key words: potato virus Y; NTN strain; coat protein; monoclonal antibody;
polyclonal antibody; ELISA; immunoblot analysis; amino acid sequencing
Acta virologica 43: 391 - 393, 1999
J. BLAHOVÁ, K. KRÁLIKOVÁ, V. KRČMÉRY, SR., P. JEŽEK
Institute of Preventive and Clinical Medicine, Limbová 14, 833 01 Bratislava, Slovak
Republic;
Municipal Hospital, Příbram, Czech Republic
Summary. - Isolation and propagation of a wild type phage, isolate AP-423, from an apparently lysogenic strain of Pseudomonas aeruginosa, resistant to a series of anti-pseudomonadal antibiotics, and its use for transduction of resistance determinants is described. The phage isolate AP-423 showed a phenomenon of host restriction, i.e. it was lysogenic only for some of the recipient strains tested. Its transduction capacity, both in sets of genes transduced and frequency of transduction, was different in two recipient strains of P. aeruginosa. This phage showed also some restriction in titers, to which it could be propagated, only in certain recipient strains.
Key words: Pseudomonas aeruginosa; antibiotic resistance; transduction
Acta virologica 43: 395 - 398, 1999
M. BYSTRICKÁ, M. ZAŤOVIČOVÁ, M. PETRÍKOVÁ, Ľ. SOLÁRIKOVÁ, G. RUSS, T. ZIEGLER
Institute of Virology, Slovak Academy of Sciences, Dúbravská cesta 9, 842 45
Bratislava, Slovak Republic;
Department of Virology, University of Turku, 20520 Turku, Finland;
Department of Medical Microbiology, University of Oulu, 90220 Oulu, Finland
Summary. - Eight monoclonal antibodies (MAbs) to herpes simplex viruses 1 and/or 2 (HSV-1, HSV-2)) were tested for their reactivity with 190 previously typed HSV-positive clinical specimens in order to determine their suitability for use as type-specific reagents. Using a rapid culture assay we found that two MAbs (T51 and T96) identified HSV-1 in all the 94 specimens, previously found HSV-1-positive, but did not react with the 96 specimens, previously found HSV-2-positive. In contrast, one MAb (303) confirmed the presence of HSV-2 in all the specimens, previously found HSV-2-positive, but did not react with any of the 94 specimens, previously found HSV-1-positive. These three type-specific MAbs allow for a rapid type-specific identification of HSV in infected cultures. One type-common MAb (T111) reacted with all HSV-positive cultures. This MAb can be used as an excellent reagent for detection of HSV infection.
Key words: herpes simplex virus; typing; clinical specimen; monoclonal antibody;
rapid culture assay
Acta virologica 43: 399 - 402, 1999
I. Davidson, Y. Weisman
Division of Avian and Fish Diseases, Kimron Veterinary Institute, Bet Dagan, P.O. Box 12, 50250 Israel
Summary. - Two new turkey meningoencephalitis virus (TMEV) nucleotide sequences were aligned to complete sequences of genomes of the flaviviruses that were available at present in the GeneBank. It was found that the both TMEV sequences represent different NS5 locations; the sequence with Acc. No. AF098456 is located downstream of that with Acc. No. AF013377 on the TMEV NS5 gene. This finding provides further insight into the TMEV NS5 gene structure and shows that the two sequences are located on the NS5 gene separately.
Key words: turkey meningoencephalitis virus; NS5 gene; alignment; nucleotide
sequence
Acta virologica 43: 403 - 405, 1999