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Articles by C. Ding
Total Records ( 2 ) for C. Ding
  H Chen , W Wang , C Song , S Yu and C. Ding
 

Currently available vaccines against Mycobacterium bovis, the causative agent of tuberculosis, do not provide reliable efficacy and there is therefore a need for a novel vaccine with improved efficacy. Here, we use protein transduction technology to deliver DNA vaccines expressing mycobacterial antigens directly to target cells. We used various protein transduction domain (PTD) proteins including the VP22 conjugate from Marek's disease virus serotype 1 (MDV-1), as delivery systems for DNA constructs encoding the antigens early secretory antigenic target-6 kDa (ESAT-6) protein and culture filtrate protein 10 (CFP-10) of M. bovis. The eukaryotic expression plasmid pZ106, encoding antigens ESAT-6 and CFP-10, conjugated to various PTDs, was used to construct experimental preparations. Our findings demonstrated that VP22 alone or in combination with CFP-10:ESAT-6 fusion protein could spread into all the nuclei of the cell monolayer surrounding the transfected cells. Whereas trans-activating transcriptional PTD showed limited delivery of the fusion protein and 8R peptide was unable to deliver the fusion protein into any untransfected cells. We have demonstrated that immunization with a preparation fused to VP22 leads to a higher antibody and interferon- titer (P < 0.05). Taken together, our results demonstrated that MDV-1 VP22 serves as a potential immune enhancer in gene therapy and immunization using DNA vaccines, offering a novel approach for the prevention of M. bovis infection.

  J Luan , J Yuan , X Li , S Jin , L Yu , M Liao , H Zhang , C Xu , Q He , B Wen , X Zhong , X Chen , H. L.Y Chan , J. J.Y Sung , B Zhou and C. Ding
 

Background: Variations in the hepatitis B virus (HBV) genome may develop spontaneously or under selective pressure from antiviral therapy. Such variations may confer drug resistance or affect virus replication capacity, resulting in failure of antiviral therapy.

Methods: A duplex PCR was used to amplify the region of the reverse transcriptase gene, the precore promoter, and the basal core promoter of the HBV genome. Four multiplex primer-extension reactions were used to interrogate 60 frequently observed HBV variants during antiviral therapy. Automated MALDI-TOF mass spectrometry (MS) was used for mutation detection. Capillary sequencing was used to confirm the MS results.

Results: The limit of quantification was 1000 HBV copies/mL for multiplex detection of HBV variants. Fifty-three variants (88.3%) were analyzed successfully in at least 90% of the sera from 88 treatment-naive patients and 80 patients with virologic breakthrough. MS was able to detect twice as many minor variants as direct sequencing while achieving close to full automation. MS and direct sequencing showed only 0.1% discordance in variant calls.

Conclusions: This platform based on multiplex primer extension and MALDI-TOF MS was able to detect 60 HBV variants in 4 multiplex reactions with accuracy and low detection limits.

 
 
 
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