Publications (5)13.26 Total impact
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Article: A single amino acid V4I substitution in VP1 attenuates virulence of very virulent infectious bursal disease virus (vvIBDV) in SPF chickens and increases replication in CEF cells.
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ABSTRACT: Infectious bursal disease virus (IBDV) is a birnavirus that causes immunosuppressive disease in chickens. The emergence of very virulent IBDV (vvIBDV) has brought new challenges for this disease. The molecular determinants for the high pathogenicity of vvIBDV are not fully understood. Previous studies focused mostly on the VP2 protein on segment A, but recent evidence suggests that segment B also plays an important role. Previously we identified eight amino acid changes in the VP1 protein of vvIBDV. In this study, we investigated effect of amino acids substitutions in VP1 on viral replication and pathogenicity. We identified a Valine to Isoleucine substitution at amino acid position 4 (V4I) of VP1 that attenuates viral pathogenicity and reduces viral replication in SPF chickens but increases viral replication in CEF cells. This study confirms that VP1 of segment B play an important role in viral replication and pathogenicity of vvIBDV.Virology 03/2013; · 3.35 Impact Factor -
Article: A simple and efficient method to rescue very virulent infectious bursal disease virus using SPF chickens.
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ABSTRACT: Reverse genetic systems for efficient generation of very virulent infectious bursal disease virus (vvIBDV) are currently limited. In this study, we have developed a simple and efficient way to rescue vvIBDV using SPF chickens. The genome of a vvIBDV strain, HLJ0504, flanked by hammerhead and hepatitis delta ribozyme sequences, was cloned downstream of the cytomegalovirus enhancer and the chicken beta-actin promoter of the vector pCAGGS. After transfection of DF-1 cells, cell suspensions were injected into the bursa organ of three-week-old SPF chickens. Using this system, vvIBDV was recovered at high titers after one passage, and the rescued vvIBDV remained highly lethal to SPF chickens. This simple and efficient method to rescue vvIBDV will be a valuable tool for better understanding the molecular virulence determinants of vvIBDV.Archives of Virology 02/2012; 157(5):969-73. · 2.11 Impact Factor -
Article: A one-step reverse transcription loop-mediated isothermal amplification for detection and discrimination of infectious bursal disease virus.
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ABSTRACT: Infectious bursal disease (IBD) is a highly contagious immunosuppressive disease in young chickens caused by infectious bursal disease virus (IBDV). It causes huge economic losses to the poultry industry. The objective of this study is to develop a loop-mediated isothermal amplification (LAMP) method for the detection and discrimination of IBDV. In this study, we applied reverse transcription loop-mediated isothermal amplification (RT-LAMP) to detect IBDV in one simple step and further identified the very virulent strain from non-vvIBDVs with a simply post-amplification restriction enzyme analysis. Based on sequence analysis, a set of two inner, two outer and two loop primers were designed to target the VP5 gene and they showed great specificity with no cross reaction to the other common avian pathogens. The detection limit determined by both color change inspection and agarose gel electrophoresis was 28 copies viral RNA, which was almost as sensitive as a real-time RT-PCR previous developed in our laboratory. We also identified a unique Tfi I restriction site located exclusively in non-vvIBDVs, so very virulent strain could be distinguished from current vaccine strains. By screening a panel of clinical specimens, results showed that this method is high feasible in clinical settings, and it obtained results 100% correlated with real-time RT-PCR. RT-LAMP is a rapid, simple and sensitive assay. In combination with the Tfi I restriction analysis, this method holds great promises not only in laboratory detection and discrimination of IBDV but also in large scale field and clinical studies.Virology Journal 03/2011; 8:108. · 2.34 Impact Factor -
Article: Molecular characteristics of segment B of seven very virulent infectious bursal disease viruses isolated in China.
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ABSTRACT: Infectious bursal disease virus (IBDV) is a birnavirus that causes immunosuppressive disease in chickens. Segment B of IBDV encodes the RNA-dependent RNA polymerase VP1, which is involved in virulence. We sequenced and analyzed segment B from seven Chinese IBDV isolates, all belonging to very virulent IBDV (vvIBDV), and clustered into Branches II and III. Phylogenetic analysis suggested that segment B of the HLJ isolates in Branch II might have originated from an unidentified host, and HuB-1 might have originated in Europe. Eight aa (4V, 61I, 145T, 287A, 508K, 511S, 646S, and 687P) were conserved in Branches II and III, and may contain potential segment B virulence determinants. Five aa (146D, 242E, 390M, 562P, and 695R) were found only in Branch III, and may be origin characteristics. Moreover, 55T and 63A in the 5'-untranslated region (UTR), and 2786C in the 3'-UTR were conserved in vvIBDV and may function in UTR secondary structure.Virus Genes 10/2010; 41(2):246-9. · 1.85 Impact Factor -
Article: A single amino acid in the C-terminus of VP3 protein influences the replication of attenuated infectious bursal disease virus in vitro and in vivo.
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ABSTRACT: The very virulent infectious bursal disease virus (vvIBDV) Gx strain causes over 60% mortality in chickens but cannot replicate in CEF cultures. The attenuated Gt strain, however, is not virulent in chickens and replicates well in CEF cultures. The two strains display differences in 6 amino acids in VP4 and 4 amino acids in VP3. To determine whether VP4 and VP3 are involved in the virulence and replication of IBDV, three chimeric viruses, in which the VP4/VP3/3'UTR, VP3/3'UTR or VP4 region of Gt were replaced by the corresponding region of Gx, were constructed and characterized in vitro and in vivo. The substituted regions in VP4 or VP3 did not affect virulence of Gt. While the substituted region in VP4 had no effect on viral replication of Gt in CEF cultures, substitution of the VP3/3'UTR region did reduce the replicative capacity of the virus. Through site-directed mutagenesis, three rescued recombinant viruses with a single amino acid substitution in the C-terminus of VP3 of the Gt strain (L981P, A990V and T1005A) were characterized in a similar manner. Amino acid substitution at position 990 reduced viral replication of Gt and reduced its efficacy of protection against vvIBDV Gx challenge in vivo. This study provides important information for the design and development of more effective IBDV vaccines using reverse genetics.Antiviral research 08/2010; 87(2):223-9. · 3.61 Impact Factor
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- Virology (1)
- Archives of Virology (1)
- Virus Genes (1)
- Virology Journal (1)
- Antiviral research (1)
Institutions
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2010–2013
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Harbin Veterinary Research Institute
Harbin, Heilongjiang Sheng, China
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