ORIGINAL_ARTICLE
Development of an ELISA Test for Detection of Anti FMD Virus Type A87/IRN Antibody Using Irradiated Antigen
Background and Aims: The aim of this study was application of gamma irradiated FMDV antigen for liquid phase blocking sandwich ELISA (B-ELISA) to quantify guinea pig anti FMDV type A87/IRN antibody. Methods: FMDV type A87/IRN was propagated to a titer of 107.5/ml TCID50 and then irradiated by gamma irradiator. The irradiated 146 S antigen was purified by sucrose gradient centrifugation after inactivation, and used for inoculation of rabbits and guinea pigs. The anti-FMDV sera were collected from the animals. ELISA test was optimized and the titer of antibodies was compared with that of virus neutralization test (VNT) to determine the correlation between these techniques. Results: The results of VNT and B-ELISA did not show significant differences (P>0.05). The sensitivity and specificity of B-ELISA in comparison to VNT for detection of anti FMDV type A 87/IRN antibody were calculated to be %94.73 and %90.9 respectively. Conclusion: The ELISA titers in sera collected from animals were higher than those recorded by VNT. These results suggested that, in addition to neutralizing antibodies, the ELISA was measuring other classes of antibodies which did not neutralize FMDV in vitro. Also comparison of the VNT and B- ELISA results by t-test showed that the gamma irradiated inactivated FMDV antigen has unaltered antigenicity.
http://journal.isv.org.ir/article-1-37-en.pdf
2014-10-12
1
6
10.21859/isv.3.3.1
Foot and Mouth Disease Virus
ELISA
antibody
sera
F
Motamedi-Sedeh
fmotamedi@nrcam.org
1
Agricultural, Medical and Industrial Research School; Nuclear Science & Technology Research Institute, Karaj, Iran
AUTHOR
A
Khorasani
2
Razi vaccine and serum research Institute, Karaj, Iran
AUTHOR
H
Mahravani
3
Agricultural, Medical and Industrial Research School; Nuclear Science & Technology Research Institute, Karaj, Iran
AUTHOR
K
Shafae
4
Agricultural, Medical and Industrial Research School; Nuclear Science & Technology Research Institute, Karaj, Iran
AUTHOR
ORIGINAL_ARTICLE
Genetic Analysis of Hemagglutinin Protein of H9N2 Isolated from Live Bird Markets in Tehran Province in 2007-2008
Background and Aims: Influenza A virus subtype H9N2 have continued to circulate in domestic poultry farms in Asia since 1998. The virus circulates in live bird markets, missing link in epidemiology of avian influenza. Regarding previous studies on H9N2 viruses of Iran and having no data on this subject in Iranian live bird markets this study was conducted to analyze genetically hemagglutinin protein of H9N1 virus. Methods: A total of 500 tracheal and cloacal swabs from clinically healthy birds of Tehran's live bird markets were collected. Diagnostic RT-PCR was done on them using specific primers for subtype H9N2. Eight positive samples were selected for inoculation into 9-11 days SPF emberyonated eggs and the virus was grown and isolated. Amplification of the HA gene was carried out by PCR using two pairs of specific primers. PCR products were separated, purified and cloned. The products were sequenced and analyzed with M13 primers. They shared high amino acid homology with genes of other H9N2 viruses isolated previously in Asia and Iran. Results: H9N2 viruses isolated from live bird markets were highly similar to viruses isolated from industrial poultry being circulated as early as 2001. Conclusion: The results suggest that a common source of H9N2 viruses is circulating in Iran.
http://journal.isv.org.ir/article-1-38-en.pdf
2014-10-12
7
15
10.21859/isv.3.3.7
Avian influenza virus (H9N2)
Live Bird Market
Hemagglutinin
S
Ghadi
1
Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
AUTHOR
MH
Bozorgmehri-Fard
mhbfard@ut.ac.ir
2
Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
AUTHOR
V
Karimi
3
Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
AUTHOR
M
Tavassoti-Kheiri
4
Influenza Unit, Pasteur Institute of Iran, Tehran, Iran
AUTHOR
MH
Rafiei
5
Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
AUTHOR
ORIGINAL_ARTICLE
Cloning & Expression of F Protein Gene (HR1 region) of Newcastle Disease Virus NR43 Isolate from Iran in E.coli
Background and Aims: NDV (Newcastle Disease Virus) is one of the viruses that cause disease in avian with severe economic losses in the poultry industry in many countries. Fusion protein (F) which plays a major role in the virus pathogenicity contains several regions that have a role in the fusion process. Mutation in the sequence of HR1 & HR2 regions of this protein prevents fusion of the virus to host cell. In addition, the proteins of HR1 and HR2 regions have antitumor properties that are related to their pathogenicity. Methods: In this investigation we used Newcastle disease virus NR43 isolate, from poultry diseases diagnostic department of Razi vaccine and serum research institute. RNA was extracted using SDS and proteinase K procedure. In the next step, RT-PCR was carried out and then cDNA cloned in pTZ57RT vector. After sequencing and alignmenting of the cDNA, a pair of proper primers for cloning HR1 in expression vector Pet32a(+) was designed. The HR1 expression was carried out by SDS–PAGE Western- Blotting in which the peptides were blotted onto nitrocellulose membrane using Ni-NTA anti His tag (1:1500 dilutions) coupled to HRP enzyme. Results: A peptide with 23.76 kD/a molecular weight s peptide was obtained. Conclusion: By cloning and expression of HR1 region of protein F, it will be possible to express the whole gene that could be introduced as a novel vaccine against NDV.
http://journal.isv.org.ir/article-1-39-en.pdf
2014-10-12
16
22
10.21859/isv.3.3.16
Cloning
Newcastle Disease Virus (NDV)
F protein gene
Heptad repeat
M
Frozandeh-Moghadam
1
Department of Medical biotechnology, Faculty of Medical Sciences, Tarbiat Modares University. Tehran, Iran
AUTHOR
R
Madani
mr_madani@yahoo.com
2
Department of Biotechnology, Razi Vaccine and Serum Research Institute, Karaj, Iran
AUTHOR
MR
Dehghani
3
Department of Medical biotechnology, Faculty of Medical Sciences, Tarbiat Modares University. Tehran, Iran
AUTHOR
SL
Mosavi
4
Department of Biology, Imam Hossein University, Tehran, Iran
AUTHOR
SA
Pourbakhsh
5
Department of Research & Diagnosis of Poultry Disease, Razi Vaccine and Serum Research Institute, Tehran, Iran
AUTHOR
F
Golchinfar
6
Department of Biotechnology, Razi Vaccine and Serum Research Institute, Karaj, Iran
AUTHOR
ORIGINAL_ARTICLE
Poliovirus Particles do not Form in Preinfected Cells with Reovirus
Background and Aims: Inhibition of viral growth in coinfected cells with two different viruses has been described. This phenomenon known as viral interference can occur in several virus host systems such as interference of enterovirus infection on poliovirus vaccine strains. In this study we superinfected reovirus infected HeLa cells with poliovirus to determine if poliovirus can replicate in such cells and form mature virus particles. Methods: Cells were infected first with reovirus then were reinfected with poliovirus. The amounts of viral particles were measured by electron microscopy and plaque assay titration. The amount of viral yield was also measured using the technique of real time RT-PCR for measuring the viral load in infected cells. Results: In cells infected first with reovirus and then superinfected with polio virus, only reovirus particles were produced. Virus production was determined by assaying viral titer using the plaque assay technique and electron microscopy. There was no poliovirus particles observable in the superinfected cells. The amount of poliovirus load in reovirus infected cells was also drastically reduced. Conclusion: The growth of poliovirus was inhibited in reovirus infected cells and no infectious poliovirus particles could be observed. This observation could be important to consider in poliovirus vaccination program.
http://journal.isv.org.ir/article-1-40-en.pdf
2014-10-12
23
28
10.21859/isv.3.3.23
Poliovirus
Reovirus
Interference
Coinfection
A
Ataei-Pirkooh
1
Department of Virology, Faculty of Medicine, Iran university of Medical Sciences, Tehran, Iran
AUTHOR
M
Shamsi-Shahrabadi
2
Department of Virology, Faculty of Medicine, Iran university of Medical Sciences, Tehran, Iran
AUTHOR
HR
Monavari
3
Department of Virology and Antimicrobial Resistance Research Center, Iran University of Medical Sciences, Tehran, Iran
AUTHOR
A
Shafiei
4
Razi Vaccine and Serum Institute, Karaj, Iran
AUTHOR
ORIGINAL_ARTICLE
Construction of the Chimeric HSP70 - E7 Vector and Evaluation of its Protein Production
Background and Aims: Since the produced recombinant proteins by molecular genetics techniques commonly have some limitations in the application, chimeric protein are introduced. Chimeric proteins have found widespread application for the study of protein folding, structure stability, function and immunogenisity. Methods: According to the known Immunomodulatory effect and structure of HSP70 molecule, full length human HSP70 was selected and a hybrid was made with HPV-E7gene.The recombinant plasmid pCDNA3.1/ E7-HSP70 was constructed using sequential PCR and cloning steps. The entire upstream and downstream sequences of the target molecules were synthesized separately. The sequential cloning was performed for cloning of the entire sequences of the target molecule of HPV16-E7 fragment in pcDNA3.1. Target DNA was visualized by staining with ethidium bromide. The Cos-7 cell lines were transfected with fusion proteins in 6 well microtiter plates. After 48 hours of transfection, the target cells were removed and to SDS-PAGE analysis for mRNA detection. Immunoreactivity of the protein product was assayed by Western blotting using monoclonal antibody. Results: The sequencing analysis showed that E7 gene was fused in frame to the HSP70 gene in pcDNA3.1/E7-HSP70. Western blot analysis of recombinant fusion protein using HSP 70 monoclonal antibody showed desired band as expected. The chimeric structure was expressed in cells, as expected. The resulting E7- HSP70 fusion gene would be a useful construct for future research. Conclusion: In order to change and enhance of the tropism and immunogenicity of recombinant protein, chimeric E7- HSP70 Vector was constructed. Since monovalent molecule and vaccines were clinically ineffective or poorly immunogenic, so applications of covalently linked product are introduced. The successful expression of the E7- Hsp70 fusion protein can be used as a molecular target for establishment of DNA and recombinant protein vaccine in future research.
http://journal.isv.org.ir/article-1-41-en.pdf
2014-10-12
29
34
10.21859/isv.3.3.29
pcDNA3.1
HSP70
chimeric vector
H
Razavi-Nikoo
1
Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
AUTHOR
H
Soleimanjahi
soleim_h@modares.ac.ir
2
Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
AUTHOR
F
Fotouhi
3
Influenza Unit Pasteur Institute of Iran, Tehran, Iran
AUTHOR
A
Ghaemi
4
Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
AUTHOR
M
Fazeli
fazelim9@gmail.com
5
Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
AUTHOR
ORIGINAL_ARTICLE
Assessment of Serum Neutralizing Antibody Titers against Foot-and-Mouth Disease Virus A and O in Young Colostrum-Fed Dairy Calves
http://journal.isv.org.ir/article-1-42-en.pdf
2014-10-12
35
38
10.21859/isv.3.3.35
Serum Neutralizing Antibody
Foot and Mouth disease
MH
Chahe
1
Animal Viral Vaccines Department, Razi Vaccine and Serum Research Institute, Karaj, Iran
AUTHOR
H
Mahravani-Behbahani
mahravani@rvsri.ir
2
Foot-and-mouth Disease Vaccine Department, Razi Vaccine and Serum Research Institute, Karaj, Iran
AUTHOR
B
Alirezaie
3
Human Viral Vaccines Department, Razi Vaccine and Serum Research Institute, Karaj, Iran
AUTHOR
G
Afshari
4
Faculty of Veterinary Medicine, Tehran University, Tehran, Iran
AUTHOR
ORIGINAL_ARTICLE
Sequence Analysis of Expressed cDNA of Bean Common Mosaic Virus RU1 Isolate
http://journal.isv.org.ir/article-1-43-en.pdf
2014-10-12
39
41
10.21859/isv.3.3.39
Bean Common Mosaic Virus
RU1
M
Naderpour
m.naderpour@dias.kvl.dk
1
Seed and Plant Certification Research Institute, P. O. Box 31535-1516, Karaj, Iran. Tel/fax: (+98) 261 27 40 777
AUTHOR
OS
Lund
2
Department of Plant Biology and Biotechnology, Faculty of Life Sciences, Copenhagen University, Copenhagen, Denmark
AUTHOR
IE
Johansen
3
Department of Genetics and Biotechnology, Faculty of Agricultural Sciences, University of Aarhus, Aarhus, Denmark
AUTHOR