1.Genetic Variation Analyses of nsp2 Gene of PRRSV in Ningxia Hui Autonomous Region of China
Hong TIAN ; Jingyan WU ; Shuanghui YIN ; Youjun SHANG ; Ziping MAN ; Na ZHAO ; Ye JIN ; Xiangtao LIU
Virologica Sinica 2009;24(3):221-226
To gain a better understanding of the genetic diversity and evolution of PRRSV in the Ningxia Hui Nationality Autonomous Region (Ningxia) of China, the nsp2 genes from a series of PRRSV strains collected from the region in 2007 were partially sequenced. These sequences were then analyzed along with the classical strain (ch-la) and two other epidemic strains SD (3) and SD2006. Comparison of the nucleotide sequence with ch-la indicated that nsp2 genes of seventeen Ningxia isolates (NX strain) have deletions of 87 nucleotides. Sequence analysis indicated that homology between the Ningxia strain and ch-la was 60.3%-79.9% in the nucleotide sequence, and homology between the NX strains and SD strains was 80.3%-98.8% in the nucleotide sequence. The nsp2 genes of the seventeen isolates had 74.9%-100% nucleotide sequence identities with each other. This study was undertaken to assess the regional variation of prevalent PRRSV and to establish a sequence database for PRRSV molecular epidemiological studies.
2.An ELISA Based on a Truncated Soluble ORF2 Protein for the Detection of PCV2 Antibodies in Domestic Pigs
Shuanghui YIN ; Shunli YANG ; Hong TIAN ; Jinyan WU ; Youjun SHANG ; Xuepeng CAI ; Xiangtao LIU
Virologica Sinica 2010;25(3):191-198
Postweaning multisystemie wasting syndrome (PMWS) is an important swine disease that is closely associated with porcine circovirus type 2 (PCV2). The capsid protein (Cap protein) is a major structural protein that has at least three immunoreactive regions, and it can be a suitable candidate antigen for detecting the specific antibodies of a PCV2 infection. In the present study, an indirect enzyme-linked immunosorbent assay (TcELISA)based on a truncated soluble Cap protein produced in Escherichia coli (E.coli) was established and validated for the diagnostic PCV2 antibodies in swine. The TcELISA was validated by comparison with an indirect immunofluorescence assay (IIFA). The diagnostic sensitivity (DSN), specificity (DSP), and accuracy of the TcELISA were 88.6%, 90.7% and 89.4%, respectively. The agreement rate was 89.38% between results obtained with TcELISA and IIFA on 113 field sera. A cross-reactivity assay showed that the method was PCV2-specific by comparison with other sera of viral disease. Therefore ,the TcELISA will be helpful for the development of a reliable serology diagnostic test for large scale detection of PCV2 antibodies and for the evaluation of vaccine against PCV2 in swine.
3.Establishment and application of a solid-phase blocking ELISA method for detection of antibodies against classical swine fever virus
Yuying CAO ; Li YUAN ; Shunli YANG ; Youjun SHANG ; Bin YANG ; Zhizhong JING ; Huichen GUO ; Shuanghui YIN
Journal of Veterinary Science 2022;23(5):e32-
Background:
Classical swine fever (CSF) is a severe infectious disease of pigs that causes significant economic losses to the swine industry.
Objectives:
This study developed a solid-phase blocking enzyme-linked immunosorbent assay (spbELISA) method for the specific detection of antibodies against the CSF virus (CSFV) in porcine serum samples.
Methods:
A spbELISA method was developed based on the recombinant E2 expressed in Escherichia coli. The specificity of this established spbELISA method was evaluated using reference serum samples positive for antibodies against other common infectious diseases.The stability and sensitivity were evaluated using an accelerated thermostability test.
Results:
The spbELISA successfully detected the antibody levels in swine vaccinated with the C-strain of CSFV. In addition, the detection ability of spbELISA for CSFV antibodies was compared with that of other commercial ELISA kits and validated using an indirect immunofluorescence assay. The results suggested that the spbELISA provides an alternative, stable, and rapid serological detection method suitable for the large-scale screening of CSFV serum antibodies.
Conclusions
The spbELISA has practical applications in assessing the vaccination status of large pig herds.