Electronic Library of Scientific Literature
Volume 42 / February 1998 / number 1
S. PARANJPE, K. BANERJEE
National Institute of Virology, P.O. Box 11, 20-A, Dr. Ambedkar Road, Pune 411 001, India
Summary. - A simple and rapid reverse transcription/polymerase chain reaction (RT-PCR) assay for detection of Japanese encephalitis virus (JEV) envelope (E) gene sequences in various biological samples is described. The assay successfully amplified JEV E gene sequences from infected cell cultures, Aedes aegypti larvae, mosquitoes and mouse blood. The sensitivity of the assay was currently 1 ng of JEV RNA and could be increased up to 1 pg on the background of 1 µg of cellular RNA by biotinylation of the PCR product, Southern blot analysis and streptavidin/alkaline phosphatase detection.
Key words: Japanese encephalitis virus; envelope gene; reverse
transcription; polymerase chain reaction
Acta virologica 42: 5 - 11, 1998
X. LÉRY, S. ABOL-ELA, J. GIANNOTTI
Entomovirology Laboratory, ORSTOM, B.O. 26, Giza code 12211, Cairo, Egypt
Summary. - Genetic heterogeneity of a wild-type granulovirus (Tunisia isolate) of the potato tuber moth Phthorimaea operculella (Phthorimaea operculella granulovirus, Phop GV) has been studied. The heterogeneity was indicated by the presence of several submolar fragments in the profiles obtained by use of several restriction endonucleases. It was also demonstrated by variations in the restriction profile of the wild-type Tunisia isolate that had underwent since 1991 in our laboratory numerous passages in vivo. A comparison of the Tunisia isolate used in Egypt in the biological control programme with other PhopGV isolates indicated that it could not be related to any of the 3 genotypes previously defined. Five clones obtained from the Tunisia isolate in vitro were further grown both in vitro and in vivo. The restriction analysis of these clones demonstrated that none of them was identical to the parental wild type virus and to any other PhopGV geographic isolates. Genotypic differences between the clones were also shown. A 19 kbp BamHI fragment absent in the original Tunisia isolate but present in its passages since 1995 at a submolar concentration, was always present at a molar concentration in its clones. The presence of this fragment reflects probably a selection of one or more variants present in the original isolate and its possible adaptation to the growth in our laboratory conditions.
Key words: potato tuber moth; Phthorimaea operculella;
granulovirus; virus clones; cell line; genetic virus heterogeneity
Acta virologica 42: 13 - 21, 1998
M. POLJAK, J. TOMAŽIČ, K. SEME, M. MATIČIČ, L. VIDMAR
Slovenian AIDS Reference Centre, Institute of Microbiology and Immunology,
Medical Faculty of Ljubljana, Zaloška 4, 1105 Ljubljana;
Department of Infectious Diseases, Clinical Centre, Japljeva 2, 1000 Ljubljana, Slovenia
Summary. - A 32 bp deletion in the CCR5 gene designated CCR5delta32 has been identified recently as the cellular basis for resistance to human immunodeficiency virus type 1 (HIV-1) in some individuals which remained non-infected despite a repeated exposure to this virus. The prevalence of this deletion was examined by polymerase chain reaction (PCR) on 51 HIV-1-infected and 385 non-infected individuals from all parts of Slovenia. 84.4% of the the HIV-1-infected and 83.2% of the non-infected individuals were homozygous for wild type CCR5, and 19.6% and 16.3%, respectively, were heterozygous. No homozygous mutant genotype was observed among the HIV-1-infected patients. Of the non-infected individuals, 2 women (0.5%) were found to harbour the CCR5delta32/CCR5delta32 genotype only, which is, to the best of our knowledge, the lowest prevalence of this particular genotype found among Caucasians to date.
Key words: human immunodeficiency virus type 1; acquired immunodeficiency
syndrome; CCR5 gene; chemokine receptor; Slovenia
Acta virologica 42: 23 - 26, 1998
E.A. SANTOS, C. NIEL, C.O.A. VIANNA, C.A. MORAES DE SÁ, S.A. GOMES
Department of Virology, Oswaldo Cruz Institute, Avenida Brasil 4365,
21045-900 Rio de Janeiro;
Gaffrée and Guinle University Hospital, Rio de Janeiro, Brazil
Summary. - Serum samples from 56 human immunodeficiency virus type 1 (HIV-1)-infected adult men were analysed for the presence of hepatitis B virus (HBV) serological markers. Two or more samples from each patient, collected over an interval of 1-6 years, were tested. The antibody against HBV core antigen (anti-HBc) prevalence was 79%. Three (5%) patients No. 5, 7, and 9 were chronic carriers of HBV surface antigen (HBsAg). HBV DNAs from serial samples of these three patients and from two HIV-seronegative control patients were characterised after amplification of different genome regions by polymerase chain reaction (PCR). Size and restriction analyses of the PCR products showed that samples from patients No. 7 (with chronic active hepatitis) and 9 (asymptomatic) contained heterogeneous HBV DNA populations. In patient No. 7, HBV DNA contained a precore gene stop codon mutation at nucleotide (nt) 1896. In addition, a deletion in the core gene was found in a sample collected two years after the onset of acquired immunodeficiency syndrome (AIDS). PCR products from serial samples of patient No. 9 indicated a mixture of HBV DNA molecules that were cloned. Sequencing of the pre-S region of the clones and phylogenetic analysis showed that patient No. 9 was superinfected with three HBV populations of distinct origin, all belonging to genotype A. HBV DNA of patient No. 5 (with AIDS) did not present any variability during a 6-year follow-up. Although two of three HIV/HBV coinfected patients harboured heterogeneous HBV DNA populations during the follow-up, no common event with respect to HBV DNA evolution was observed among the coinfected patients.
Key words: hepatitis B virus; human immunodeficiency virus; coinfection;
mutation; polymerase chain reaction; nucleotide sequencing; restriction
analysis; molecular cloning
Acta virologica 42: 27 - 33, 1998
B. PANDEY, A. ICHINOSE, A. IGARASHI
Department of Virology,
Central Laboratory, Institute for Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki City 852-8523, Japan
Summary. - Morphological changes such as loss of cytoplasm, membrane destruction and vacuolar swelling in Aedes albopictus clone C6/36 cells infected with dengue virus 2 (DV-2) New Guinea B strain and incubated at 28°C and 37°C, were observed under electron microscope. Both infected and uninfected cells showed significant damage at 37°C in contrast to normal appearance at 28°C. A higher number of virus particles were observed in the cytoplasm at 37°C than at 28°C.
Key words: dengue-2 virus; mosquito cells; elevated temperature;
Acta virologica 42: 35 - 39, 1998
M. KÚDELOVÁ, A. VOJVODOVÁ, J. RAJČÁNI
Institute of Virology, Slovak Academy of Sciences, Dúbravská cesta 9, 842 46 Bratislava, Slovak Republic
Summary. - Parallel sequencing of UL53 gene of four strains of herpes simplex virus type 1 (HSV-1), two of which (HSZP and ANGpath) were of the syn phenotype while another two (KOS and 17) were of the non-syn phenotype, showed in three strains amino acid mutations unrelated to the already described syn1 glycoprotein K (gK) mutations (Dolter et al., 1994). The only mutations which altered encoded amino acids were found in strains HSZP (Gln to Arg at position 198) and ANGpath (Val to Ile at position 137). Both mutations were localised outside of the two mutation clusters suspected for affecting syncytium formation. In addition, a CG/GC variation was found at positions 245-246 and 669-670. These compressions affected three codons altering amino acids (aa) 82 (Cys or Ser), 223 (Met or Ile) and 224 (Leu or Val), respectively.
Key words: herpes simplex virus type 1; strain HSZP; strain
ANGpath; syncytium formation; UL53 gene; glycoprotein K; nucleotide sequencing
Acta virologica 42: 41 - 45, 1998
M.M. HOSSAIN, H. TSUCHIE, M.A. DETORIO, H. SHIRONO, C. HARA, A. NISHIMOTO, A. SAJI, J. KOGA, N. TAKATA, J.K. MANIAR, D.G. SAPLE, K. TANIGUCHI, S. KAGEYAMA, H. ICHIMURA, T. KURIMURA
Research Institute for Microbial Diseases, Osaka University, 3-1
Yamadaoka, Suita, 565 Osaka, Japan;
JCR Pharmaceutical Co. Ltd., Kobe, Japan;
Hiroshima University Medical Hospital, Hiroshima, Japan;
Grant Medical College, Mumbai, India;
Eiken Chemical Co. Ltd., Tokyo, Japan;
Toyama Medical and Pharmaceutical University, Toyama, Japan;
Kyoto Prefectural University of Medicine, Kyoto, Japan.
Summary. - A search for gene(s) associated with anti-human immunodeficiency virus type 1 (HIV-1) activity of CD8+ T cells was attempted using molecular cloning and the relation between the anti-HIV activity of CD8+ T cells and the interleukin-9 receptor alpha chain (IL-9R-alpha) mRNA expression from the cDNA clones obtained was examined. The anti-HIV-1 activity of CD8+ T cell culture supernatants was assessed by measuring the level of HIV-1 replication in a CD4+ T cell line transfected with an infectious HIV-1 DNA clone. IL-9R-alpha mRNA was assayed by reverse transcriptase-polymerase chain reaction (RT-PCR). Of 5 cases showing high level of anti-HIV-1 activity (more than 80% suppression of HIV-1 replication), the mRNA was detected in 4 cases. Of 10 cases showing low level of anti-HIV-1 activity (less than 80% suppression of HIV-1 replication), the mRNA was detected in one case. Soluble recombinant human IL-9 receptor (rhIL-9sR) did not suppress HIV-1 replication at a concentration of 1 µg/ml. These data suggest that the IL-9R-alpha mRNA formation in CD8+ T cells may correlate with and play some role in the anti-HIV-1 activity of CD8+ T cells from HIV-1-infected individuals.
Key words: CD8+ T cells; anti-HIV-1 activity; cytokines;
Acta virologica 42: 47 - 53, 1998
E.A. PALOMBO, H.C. BUGG, R.F. BISHOP
Department of Gastroenterology and Clinical Nutrition, Royal Children's Hospital, Flemington Road, Parkville, Victoria 3052, Australia
Summary. - An atypical human rotavirus strain Z10262, isolated from a chronically infected immunodeficient child, displayed an unusual genomic RNA electrophoretic pattern. Besides, Northern blot analysis indicated that this strain contained an abnormally migrating gene 11 equivalent. Sequencing of this gene showed that it was derived from a genetic rearrangement which involved a partial duplication of the open reading frame (ORF) encoding the non-structural protein NSP5. However, the duplicated region contained a deletion and several point mutations relative to the first copy of the ORF. Phylogenetic analysis of human and animal NSP5 amino acid sequences including Z10262 revealed two groups of human proteins related to different animal proteins. The isolation and analysis of Z10262 strain provides further evidence for the genetic complexity of naturally occurring human rotaviruses.
Key words: rotavirus; non-structural protein; genetic rearrangement;
Acta virologica 42: 55 - 59, 1998
M. CHEN, M.Y. FAN, D.Z. BI, J.Z. ZHANG, Y.P. HUANG
Department of Rickettsiology, Institute of Epidemiology and Microbiology,
Chinese Academy of Preventive Medicine, P.O. Box 5, Changping 102206, Beijing,
P.R. of China;
Zhangzhou Antiepidemic Station, Zhangzhou, Fujian Province, P.R. of China
Summary. - The primers Rr 190.70p and Rr 190.602n were used to detect spotted fever group (SFG) rickettsiae by polymerase chain reaction (PCR) in ticks and small mammals collected in three different regions of China. The obtained results indicated that specific DNA fragments of SFG rickettsiae were amplified from Dermacentor silvarum, D. sinicus, D. auratus, Haemaphysalis concinna, H. wellingtoni, H. yeni, Apodemus agrarius, Microtus fortis, Clethrionomys rufocanus, Ondatra zibethica, Rattus flavipectus and hedgehog. The PCR product were digested with restriction endonucleases PstI and RsaI and the obtained electrophoretic profiles were compared with those of the prototype strains of SFG rickettsiae by the restriction fragment length polymorphism (RFLP) technique. The comparisons showed that the profiles were identical to those of Rickettsia sibirica. In addition, three new isolates of R. sibirica were obtained from H. yeni, D. sinicus and hedgehog, and designated NH-95, BJ-95 and BHJ-95, respectively. These results not only demonstrated a horizontal transmission of the rickettsiae between ticks and hosts but also suggested that R. sibirica is widely distributed in China and its hosts and vectors are various, all that indicating the existence of natural foci of North Asia tick-borne spotted fever specific to China.
Key words: Rickettsia sibirica; polymerase chain reaction;
restriction analysis; natural foci
Acta virologica 42: 61 - 64, 1998