Key Laboratory of Veterinary Biological Engineering and Technology

Nanjing, China

Key Laboratory of Veterinary Biological Engineering and Technology

Nanjing, China
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Li B.,Jiangsu Academy of Agricultural Sciences | Li B.,Key Laboratory of Veterinary Biological Engineering and Technology | Li B.,National Center for Engineering Research of Veterinary Bio products | Sun B.,Jiangsu Academy of Agricultural Sciences | And 20 more authors.
Journal of Virological Methods | Year: 2013

Hokoviruses have recently been detected as pathogens belonging to the family Parvoviridae, which comprises porcine hokovirus (PHoV) and bovine hokovirus (BHoV). In this study, we developed a loop-mediated isothermal amplification (LAMP) assay for the rapid, specific and sensitive detection of PHoV. A set of four primers specific for six regions within the PHoV VP1/2 genes was designed using online software. The reaction temperature and time were optimized at 65. °C and 60. min, respectively. LAMP products were detected by agarose gel electrophoresis or by visual inspection of a color change caused by a fluorescent dye. The method was highly specific for PHoV, and no cross-reaction was observed with porcine circovirus type 2 (PCV2), porcine parvovirus (PPV), porcine bocavirus (PBoV), pseudorabies virus (PRV), porcine reproductive and respiratory syndrome virus (PRRSV), classic swine fever virus (CSFV), or Japanese encephalitis virus (JEV). The detection limit was approximately 10 copies per reaction, which was 10 times more sensitive than conventional PCR. Furthermore, the efficiency of detection of PHoV in clinical samples was comparable to that of PCR and sequencing. These results show that the LAMP assay is a simple, rapid, sensitive and specific method for detecting PHoV. It does not require specialized equipment and can be used to detect PHoV both in the laboratory and in the field. © 2013 Elsevier B.V.


Li B.,Jiangsu Academy of Agricultural Sciences | Li B.,Key Laboratory of Veterinary Biological Engineering and Technology | Li B.,National Center for Engineering Research of Veterinary Bio products | Ma J.-J.,Jiangsu Academy of Agricultural Sciences | And 27 more authors.
Journal of Virological Methods | Year: 2012

The porcine boca-like virus (Pbo-likeV) was recently discovered in Swedish pigs with post-weaning multisystemic wasting syndrome (PMWS). In this study, a loop-mediated isothermal amplification (LAMP) assay was developed for rapid, specific and sensitive detection of Pbo-likeV. A set of four primers specific for six regions of Pbo-likeV VP1/2 genes was designed with the online software. The reaction temperature and time were optimized to 65°C and 60. min, respectively. LAMP products were detected by agarose gel electrophoresis or by visual inspection of a color change due to addition of fluorescent dye. The developed method was highly specific for detection of Pbo-likeV, and no cross-reaction was observed with other swine viruses, such as porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), porcine parvovirus (PPV) and classic swine fever virus (CSFV) found commonly in China. The lower detection limit of the LAMP assay was approximately 10 copies per reaction, and it was 100 times more sensitive than that of conventional PCR. Furthermore, the efficiency of LAMP for detection Pbo-likeV in clinical samples was comparable to PCR and sequencing. These results showed that the LAMP assay is a simple, rapid, sensitive and specific technique for detection of Pbo-likeV, and the procedure of LAMP does not rely on any special equipment. It has capacity for the detection of Pbo-likeV both in the laboratory and on farms. © 2011 Elsevier B.V.


Li W.,Jiangsu Academy of Agricultural Sciences | Li W.,Key Laboratory of Veterinary Biological Engineering and Technology | Li W.,National Center for Engineering Research of Veterinary Bio products | Mao L.,Jiangsu Academy of Agricultural Sciences | And 15 more authors.
Vaccine | Year: 2015

Classical swine fever (CSF) is a highly contagious and economically important viral disease that affects the pig industry worldwide. The glycoprotein E2 of CSFV can induce neutralizing antibodies and protective immunity, and is widely used for novel vaccine development. The objective of this study was to explore whether a tetraspanin molecule CD81 could improve the immune responses of an E2-based DNA vaccine. Plasmids pVAX-CD81, pVAX-E2 and pVAX-CD81-E2 were constructed and the expression of target proteins was confirmed in BHK-21 cells by indirect immunofluorescence assay. BALB/c mice were divided into 5 groups and immunized with different plasmids (pVAX-E2, pVAX-CD81-E2, pVAX-E2. +. pVAX-CD81, pVAX-CD81 and PBS) three times with two weeks interval. The results showed that the introduction of CD81 promoted higher humoral and cellular immune responses than E2 expression alone (. P<. 0.05). In addition, immunization with pVAX-CD81-E2 induced stronger immune responses than pVAX-E2. +. pVAX-CD81. Furthermore, four groups of pigs were immunized with pVAX-E2, pVAX-CD81-E2, pVAX-CD81 and PBS, respectively. Humoral and cellular immune responses detection showed similar results with those in mice. Compared to pVAX-E2, pVAX-CD81-E2 induced higher titers of neutralizing antibodies after viral challenge and conferred stronger protection. These results confirmed the capacity of swine CD81 enhancing the humoral and cellular responses with an adjuvant effect on CSFV DNA vaccine. This is the first report demonstrating the adjuvant effect of CD81 to enhance the DNA vaccination for swine pathogen. © 2015 Elsevier Ltd.


Yang L.,Jiangsu Academy of Agricultural Sciences | Yang L.,Key Laboratory of Veterinary Biological Engineering and Technology | Yang L.,National Center for Engineering Research of Veterinary Bio products | Li W.,Jiangsu Academy of Agricultural Sciences | And 20 more authors.
Infection, Genetics and Evolution | Year: 2016

Parainfluenza virus type 3 (PIV3) is one of the most important viral respiratory pathogens for humans and for many animals. One unique caprine PIV3 (CPIV3) strain named JS2013 was isolated in Chinese goat flocks with respiratory diseases in 2013. Now, the complete genome sequence of the strain JS2013 had been determined. A total of 15 overlapping DNA clones, covering the entire genome of the virus, were obtained by primer walking RT-PCR. The sequences of the 3' and 5' termini of the viral genome were amplified by 3' and 5' RACE. The viral genome was 15,618 nucleotides (nt) in length, which was consisted of six genes in the order 5'-leader-N-P/C/V-M-F-HN-L-tailer-3'. The junction sequences between two genes were highly conserved gene start and stop signal sequences, and trinucleotide intergenic regions (IGR) similar to those of other reported PIV3 strains. Phylogenetic analysis based on the complete genomes of JS2013 with other strains of genus Respirovirus demonstrated that the JS2013 obviously differed from HPIV1, Sendai virus, HPIV3 and other reported BPIV3 genotypes. Further analysis of HN genes of JS2013 along with two more CPIV3 strains isolated later indicated that CPIV3 strains formed a separate cluster. The results presented here suggested that CPIV3 is a new member of the genus Respirovirus. © 2015.


Wen L.,Jiangsu Academy of Agricultural Sciences | Wen L.,Key Laboratory of Veterinary Biological Engineering and Technology | Wen L.,National Center for Engineering Research of Veterinary Bio products | Wang F.,Jiangsu Academy of Agricultural Sciences | And 17 more authors.
Archives of Virology | Year: 2015

Porcine circovirus type 2 (PCV2) is the causal agent of a serious disease found in pigs. Here, we report the first detection of truncated genome sequences of PCV2 strain ZJ-R, with the genomic region encoding part of Rep and Cap with a nonviral insertion. To our knowledge, the genome of ZJ-R represents the first PCV2 DNA with a coding insertion. The PCV2 ZJ-R genome is 694 nucleotides long and has two main open reading frames. The whole genome sequence of ZJ-R may facilitate further study of the origin and evolution of PCV2. © 2014, Springer-Verlag Wien.


Bi Z.,Jiangsu Academy of Agricultural Sciences | Bi Z.,Key Laboratory of Veterinary Biological Engineering and Technology | Bi Z.,National Center for Engineering Research of Veterinary Bio products | Bi Z.,Jiangsu Electric Power Company | And 12 more authors.
Archives of Virology | Year: 2014

Canine distemper virus (CDV) infects a broad range of carnivores, including wild and domestic Canidae. The hemagglutinin gene, which encodes the attachment protein that determines viral tropism, has been widely used to determine the relationship between CDV strains of different lineages circulating worldwide. We determined the full-length H gene sequences of seven CDV field strains detected in domestic dogs in Nanjing, China. A phylogenetic analysis of the H gene sequences of CDV strains from different geographic regions and vaccine strains was performed. Four of the seven CDV strains were grouped in the same cluster of the Asia-1 lineage to which the vast majority of Chinese CDV strains belong, whereas the other three were clustered within the Asia-4 lineage, which has never been detected in China. This represents the first record of detection of strains of the Asia-4 lineage in China since this lineage was reported in Thailand in 2013. © 2014, Springer-Verlag Wien.

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