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Volume 47 / 2004 / number 1
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Articles
Short Communications
FOULONGNE, V., TURRIE`RE, C., DIAFOUKA, F., ABRAHAM, B., LASTERE, S., & SEGONDY, M.: Ganciclovir resistance mutations in ULl97 and UL54 genes of Human cytomegalovirus isolates resistant to ganciclovir 51
Letters to the Editor
UJIIE, H., SHIMIZU, T., & SHIMIZU, H.: Generalized pustular psoriasis and Hepatitis C virus infection 63
KULSHRESTHA, S., HALLAN, V., RAIKHY, G., KUMAR, A., RAM, R., GARG, I.D., & ZAIDI, A.A.: Molecular characterization of an Iris severe mosaic virus isolate from India 65
F.L. Xu1,2, Z.Q Yang1, C.C. Yang1,2*, S.Y. Xiao3, H. Xiao1, L. Wen1
*Corresponding author. Visiting professor of Institute of Virology, Medical School, Wuhan University, Wuhan, Hubei, P.R.China. E-mail: cyang@csmu.edu.tw; fax: +8864-23767469, +8627-87307966
1Institute of Virology, Medical School, Wuhan
University, 115, Dong-Hu Road, Wuhan, Hubei 430071, P.R. China;
2School of Medical Technology, Chung Shan Medical
University, 110, Section 1, Chien-Kuo North Road, Taichung, Taiwan 402,
R.O.C.;
3Departments of Pathology and Internal Medicine, and
Center for Biodefense and Emerging Infectious Diseases, University of
Texas Medical Branch, Texas, USA
Received September 2, 2003; accepted January 22, 2004
Summary. – Hantavirus HV114, isolated from urine
of a patient during epidemic of hemorrhagic fever with renal
syndrome (HFRS) in China, was subjected to a detailed serological
characterization using enzyme-linked immunosorbent assay (ELISA),
neutralization test and indirect immunofluorescence antibody assay
(IFA). It has been found that HV114 is antigenically similar to the
hantavirus A9 strain isolated in China and to the Hantaan 76-118 virus
(HTNV 76-118), but different from the hantaviruses isolated from
Apodemus agrarius in the region endemic for HFRS.
Key words: hantavirus; HV114 virus; serology; ELISA;
neutralization test; immunofluorescent assay
Acta virologica 48: 5 – 8, 2004
M.I. OLIVEIRA1*, S.P. CURTI1, C.A. FIGUEIREDO1, A.M.A. SARDINHA1, M.A.M. SALLUM2, E.L. DURIGON3
*Corresponding author. E-mail: olive40@uol.com.br; fax: +5521-30883753
1Instituto Adolfo Lutz, Sao Paulo, Brasil;
2Faculdade de Saúde Pública, Universidade de Sao Paulo,
Sao Paulo, Brasil;
3Instituto de Ciencias Biomédicas, Universidade de Sao
Paulo, Sao Paulo, Brasil
Received October 14, 2004; accepted February 18, 2004
Summary. – This study reports on molecular
analysis of a Measles virus (MV) isolate from a patient who
was infected in Japan but showed symptoms after arriving to Brazil. This
patient had typical clinical measles infection symptoms: fever, rash,
cough and coryza. After isolating the virus in B95a cells,
a fragment of the nucleoprotein (N) gene was amplified by reverse
transcription–polymerase chain reaction (RT-PCR) and subjected to
direct nucleotide sequencing. The sequence data showed that the MV
isolate of concern is of the D5 genotype.
Key words: Measles virus; nucleotide sequence; molecular
epidemiology; nucleoprotein; Brazil
Acta virologica 48: 9 – 14, 2004
M. Hosamani*, S. Nandi1, B. Mondal, R.K. Singh, T.J. Rasool, S.K. Bandyopadhyay1
*E-mail:
m_hosa@email.com; fax:
+91-5942286347
1Present address: Indian Veterinary Research Institute,
Izatanagar, Bareilly 243 122, India.
Division of Virology, Indian Veterinary Research Institute, Mukteswar, Uttaranchal 263138, India
Received October 3, 2003; accepted February 23, 2004
Summary. – An Indian isolate of Goatpox virus
(GTPV) was adapted and propagated in Vero cells for development of an
attenuated virus. The virus was initially passaged in primary lamb
testes cells and subsequently in Vero cells. At the 55th passage, the
virus showed evidence of attenuation when tested for safety in
sero-negative goats. At this stage, the virus was found to be completely
non-pathogenic. The virus was passaged further and the 60th passage was
usedfor testing its immunogenicity in goats. The latter were inoculated
with 10, 100 and 1000 TCID50 of the attenuated virus by intradermal
(i.d.) route and challenged after 28 days with virulent GTPV. The
attenuated virus produced no adverse reaction even at the highest dose
and conferred complete protection even at the lowest dose against
challenge with a high dose (2 x 106 of 50% skin-reactive
dose SRD50) of virulent virus. Increased levels of virus-specific serum
antibodies could be demonstrated by both indirect enzyme-linked
immunosorbent assay (ELISA) and virus neutralization (VN) test in all
the immunized goats. No horizontal transmission of the virus from the
immunized to in-contact animals took place. Our results suggest that
this attenuated virus could be a safe, immunogenic and potent
candidate for developing a vaccine against goatpox.
Key words: attenuated Goatpox virus; immunogenicity; Vero
cells
Acta virologica 48: 15 – 21, 2004
S.W. Tan1, A.R. Omar1*, I. Aini1, K. Yusoff2, W.S. Tan2
*Corresponding author. Fax: +603-89486317; E-mail: aro@vet.upm.edu.my
1Faculty of Veterinary Medicine,
2Faculty of Science and Environmental Studies, University
Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia
Received September 17, 2003; accepted February 25, 2004
Summary. – A two-step SYBR Green I real time
polymerase chain reaction (PCR, real time PCR) for the detection of
Newcastle disease virus (NDV) was developed. A melting curve
analysis was performed to distinguish specific from non-specific
products and primer dimers. Regardless of different virus pathotypes the
melting temperature (Tm) ranged from 86oC to 87oC. The sensitivity of
the real time PCR was compared with the reverse transcription
(RT)-nested PCR enzyme-linked immunosorbent assay (ELISA, RT-nested PCR
ELISA). Whereas the detection limit of the real time PCR was 10 pg DNA,
the RT-nested PCR ELISA and conventional PCR could only detect up to
1 ng and 10 ng DNA, respectively. Thus the real time PCR offers
a sensitive, rapid and convenient method for screening large number
of NDV specimens.
Key words: ELISA; Newcastle disease virus; SYBR Green I;
real time PCR; RT-nested PCR ELISA
Acta virologica 48: 23 – 28, 2004
Ľ. Škultéty1, L. Hernychová2, R. Toman1, M. Kroča3, J. Stulík2, A. Macela2*
*Corresponding autor. E-mail: amacela@pmfhk.cz, fax: +420 49-5513018
1Institute of Virology, Slovak Academy of Sciences, 845
05 Bratislava, Slovak Republic;
2Institute of Molecular Pathology and Proteome Centre
for the Study of Intracellular Parasitism of Bacteria, Purkynì
Military Medical Academy, 500 01 Hradec Králové, Czech Republic;
3Central Military Medical Institute, 561 66 Techonin,
Czech Republic
Received December 12, 2003; accepted March 3, 2004
Summary. – Whereas the complete genome of Coxiella
burnetii (C.b.), the etiological agent of Q-fever, has recently been
published (Seshadri et al., Proc. Natl. Acad. Sci. USA. 100, 5455–5460,
2003), the C.b. proteome is still under study. Using the bioinformatic
approach, we found in total 309 proteins on two dimensional
electroctrophoretic images of C.b. whole cell lysates. Eighteen major
protein species were subjected to peptide mass fingerprinting and
identified as the products of 6 known open reading frames (ORFs):
the chaperone DnaK (heat shock 70 K protein), chaperonin 60
K (GroEL protein, heat shock protein B), DnaJ-like protein djlA
(mucoidy activation protein mucZ), elongation factor Ts (EF-Ts),
ribosomal protein L7/L12, and chaperonin 10 K (GroES protein, heat
shock protein A).
Key words: Coxiella burnetii; proteins; peptide mass
fingerprinting; 2-D electrophoresis; MALDI-ToF
Acta virologica 48: 29 – 34, 2004
P. HUSA1*, P. SMEJKAL2, L. HUSOVÁ3, M. PENKA2
*Corresponding author. E-mail: phusa@med.muni.cz; fax: +4205-32232221
1Department of Infectious Diseases, University Hospital
Brno, Jihlavska 20, 625 00 Brno, Czech Republic;
2Department of Clinical Haematology, University Hospital
Brno, Brno, Czech Republic;
3Medical Gastroenterological Department, University
Hospital Brno, Brno, Czech Republic
Received January 16, 2004; accepted March 1, 2004
Summary. – Chronic hepatitis C infection is
common among hemophiliacs in all the developed countries. Since 1996,
only alpha-interferon (alpha-IFN) in monotherapy has been used for the
treatment of chronic hepatitis C in hemophiliacs (6 patients). In
Czech Republic a combination therapy with alpha-IFN and ribavirin
has been used since 1999 (13 patients). Finally, a combination
therapy with pegylated alpha-IFN (PEG-alpha-IFN) and ribavirin is being
used since 2001 (still 3 patients). In all cases, the treatment
lasted 48 weeks. A sustained virological response (SVR, defined as an
undetectable serum HCV RNA level 24 weeks after the treatment was
completed) was not achieved in any of 6 patients treated with
alpha-IFN alone. A combination therapy with alpha-IFN and ribavirin
yielded better results: four of eight patients still untreated with
alpha-IFN (naive patients), one of two relapsers, and one of three
non-responders to previous alpha-IFN monotherapy achieved SVR. So far
the combination therapy with PEG-alpha-IFN and ribavirin has been used
only in 3 patients. SVR was achieved in one patient who had
relapsed after the combination therapy with IFN-alpha and ribavirin, and
in 1 of 2 non-responders to this therapy. We conclude that the
efficacy and tolerability of the treatment of chronic hepatitis C in
hemophiliacs did not differ from that of chronic hepatitis C in
other patients.
Key words: chronic hepatitis C; hemophilia; alpha-interferon;
pegylated alpha-interferon; ribavirin
Acta virologica 48: 35 – 38, 2004
P. Saravanan1*, SatishKumar2, J.M. Kataria3, T.J. Rasool1
This study is a part of PhD
thesis of the first author.
*E-mail: drsaravana72@dr.com;
drsaravana72@epatra.com;
fax: +91-581-286347
1Division of Virology, Indian Veterinary Research
Institute, Mukteswar, Nainital, Uttaranchal, 263 138 India;
2National Biotechnology Centre, Indian Veterinary Research
Institute, Izatnagar, Uttar Pradesh, India;
3Division of Avian Diseases, Indian Veterinary Research
Institute, Izatnagar, Uttar Pradesh, India
Received August 12, 2003; accepted March 2,2004
Summary. – Antigenic determinant analysis was
carried out on VP3, one of major immunogenic proteins of Infectious
bursal disease virus (IBDV) using computer algorithms. Altogether 17
peptides were synthesized for predicted putative regions and were tested
for their reactivity with IBDV-positive polyclonal sera as well as with
antisera to other common avian viruses to confirm specificity and to
rule out cross reactivity. Of 17 peptides tested, three were selected
and synthesized in multiple antigenic peptide (MAP) format. The
immunization of rabbits with the three MAPs resulted in high humoral
immune response. The purified antipeptide antibodies were screened
against native IBDV antigen and the respective titers were determined.
Out of the three antisera to MAPs that raised against the MAP3, spanning
the amino acids (aa) 974–995 region on the VP3 protein had a very
high titer (2048) and reacted specifically with IBDV. Thus, the
antiserum to MAP3 detected native virus in enzyme-linked immunosorbent
assay (ELISA), revealing the presence of a potential antigenic
determinant on the C-terminus of the protein. This study proved that an
antipeptide antibody could be used as a safe and specific tool for
the diagnosis of IBD in chickens.
Key words: antipeptide antibodies; Infectious bursal disease
virus; multiple antigenic peptide; antigenic determinants; diagnosis
Acta virologica 48: 39 – 45, 2004
R. JAYAKUMAR*, K.G. TIRUMURUGAAN, G. GANGA, K. KUMANAN, A. MAHALINGA NAINAR
*E-mail: rjkumar48@yahoo.com; fax: +25362787/+253889445
Department of Animal Biotechnology, Madras Veterinary College, Tamil Nadu Veterinary and Animal Sciences University, Chennai-600 0007, India
Received October 17, 2003; accepted February 18, 2004
Summary. – Rabies occurs in all parts of Indian
sub-continent except Andaman and Nicobar and Lakshadweep group of
islands. The full-length nucleoprotein (N) gene sequence of a rabies
virus isolate from India is reported for the first time and the same has
been compared with available N gene sequences from the database. A central
domain of 230 amino acids (aa) from aa 141 to aa 370 exhibited more than
95% similarity. There were 8 amino acid positions (aa 29, 32, 38,
84, 119, 379, 438, and 439) at which substitution was unique for Indian
isolates but common for laboratory strains. In antigenic epitopes,
except for a single amino acid difference at the antigenic site IV,
the amino acids were conserved. The Indian isolate also possessed two
Bam HI sites (aa 247 and 278), while the other Asian isolates had only
one site at aa 278 or were not digested with Bam HI at all. Phylogenetic
analysis also demonstrated that the Indian isolate was closely related
to the Sri Lankan isolate and grouped in the cluster that comprised of
the isolates from other Asian countries namely China and Pakistan.
Key words: Rabies virus; Indian isolate; nucleoprotein;
nucleotide sequencing; phylogenetic analysis
Acta virologica 48: 47 – 50, 2004
V. FOULONGNE, C. TURRIERE, F. DIAFOUKA, B. ABRAHAM1, S. LASTERE2, M. SEGONDY*
*Corresponding
author. E-mail: m-segondy@chu-montpellier.fr;
fax: +33467-337793
1Present address: Département des Maladies Infectieuses et
Tropicales, Hôpital Tenon, Paris, France
2Present address: Laboratoire de Virologie, Hôpital
Bichat-Claude Bernard, Paris, France.
Laboratoire de Virologie, Hôpital Saint-Eloi, Centre Hospitalier Universitaire (CHU) de Montpellier, 80 Avenue Augustin Fliche, 34295 Montpellier Cedex 5, France
Received November 18, 2003; accepted February 24, 2004
Summary. – Human cytomegalovirus (HCMV) resistance
to ganciclovir results from mutations in viral phosphotransferase (UL97)
and/or DNA polymerase (UL54) genes. The HCMV isolates from the blood of
immunocompromised patients with persisting presence of the pp65 antigen
in the blood in spite of ganciclovir therapy were tested for ganciclovir
susceptibility by an immediate-early antigen plaque reduction assay, and
the UL54 and UL97 genes were sequenced. Nine isolates from eight
patients (six patients with acquired immune deficiency syndrome (AIDS),
one liver transplant recipient and one renal transplant recipient)
showed phenotypic resistance to ganciclovir. All these
ganciclovir-resistant HCMV isolates harbored one or more of the
following UL97 mutations: M460V, A594V, A594T, L595S, C603W, and M615V.
Two isolates harbored the P522S mutation in the UL54 gene. The M615V
mutation in the UL97 gene has not been reported earlier and its role in
ganciclovir resistance remains to be elucidated. In
ganciclovir-resistant HCMV isolates the UL54 gene was less frequently
mutated than the UL97 gene. The P522S mutation was relatively frequent
in UL54-mutated HCMV isolates.
Key words: Human cytomegalovirus; resistance; ganciclovir;
UL54; UL97
Acta virologica 48: 51 – 55, 2004
K. Kovařčík, V. Valentová*
*Corresponding author. E-mail: valentova@vri.cz; fax: +420541-211229.
Veterinary Research Institute, Hudcova 70, Brno 621 32, Czech Republic
Received January 23, 2004; accepted March 4, 2004
Summary. – In this study we showed a high
degree of genetic homogeneity among recently (2002–2003) circulating
Bovine respiratory syncytial virus (BRSV) strains in cattle population
in the Czech Republic. These strains are in a phylogenetic tree
more closely related to the Danish strains from 1995 than to the Czech
strain VS97 from 1997 that shares the highest similarity with the French
strain F1 and the Belgian strain P10. From the sequence analysis we
deduce that the revealed high diversity between BRSV strains from
2002–2003 and those from 1997, at both nucleotide (0–11.4%) and
amino acid (0–21%) level, is more likely due to distinct sources of
the virus strains than to the sequence evolution.
Key words: Bovine respiratory syncytial virus; sequence
analysis; phylogenetic analysis
Acta virologica 48: 57 – 62, 2004
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