Shao-Hui Liang

Publications (7)4.55 Total impact

• Article: Specific detection of Angiostrongylus cantonensis in the snail Achatina fulica using a loop-mediated isothermal amplification (LAMP) assay.

  Chun-Yan Liu, Hui-Qun Song, Ren-Li Zhang, Mu-Xin Chen, Min-Jun Xu, Lin Ai, Xiao-Guang Chen, Xi-Mei Zhan, Shao-Hui Liang, Zi-Guo Yuan, Rui-Qing Lin, Xing-Quan Zhu
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  ABSTRACT: Angiostrongylus cantonensis, a rat lungworm, can cause eosinophilic meningitis and angiostrongyliasis in humans following ingestion of contaminated foods or intermediate/paratenic hosts with infective larvae. The snail Achatina fulica is one of the important intermediate hosts of A. cantonensis and is commonly eaten by humans in some countries. In the present study, we developed a loop-mediated isothermal amplification (LAMP) method for the specific detection of A. cantonensis in Ac. fulica. Primers for LAMP were designed based on the first internal transcribed spacer (ITS-1) of nuclear ribosomal DNA (rDNA) of A. cantonensis. Specificity tests showed that only the products of A. cantonensis were detected when DNA samples of A. cantonensis and the heterologous control samples Anisakis simplex s.s, Trichuris trichiura, Toxocara canis, Trichinella spiralis and Ascaris lumbricoides were amplified by LAMP. Sensitivity evaluation indicated that the LAMP assay is 10 times more sensitive than the conventional polymerase chain reaction (PCR) assay. The established LAMP assay is rapid, inexpensive and easy to be performed. It can be used in clinical applications for rapid and sensitive detection of A. cantonensis in snails, which has implications for the effective control of angiostrongyliasis.
  Molecular and Cellular Probes 04/2011; 25(4):164-7. · 2.08 Impact Factor
• Article: [Microbial survey of bacterial contamination of shellfish products in Wenzhou].

  Xin Hu, Qing-yun Zhuge, Chang-wang Pan, Shao-hui Liang, Hui-cong Huang, Feng Tan
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  ABSTRACT: To investigate the status of bacterial contamination in the hellfish products in Wenzhou. One hundred samples were collected and their bacterial populations including the total plate count were investigated. Of the 100 samples collected, 67 samples failed to not meet the national regulations due to bacterial contamination, accounting for 67% of the total samples. Among the contaminated samples, the most serious contamination was caused by corliforms (61.4% of the total plate count with contamination), followed by Salmonella (18.6%), Vibio parahaemolyticus (15.7%), Listeria spp. (4.3%) and others (6%). Microbial pollution has become a threat to the marine shellfish products in Wenzhou.
  Nan fang yi ke da xue xue bao = Journal of Southern Medical University 07/2010; 30(7):1624-5.
• Article: [Expression and antigenicity analysis of NTPase gene of Toxoplasma gondii].

  Dan Sha, Feng Tan, Chang-Wang Pan, Shao-Hui Liang
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  ABSTRACT: To clone and express prokaryotic recombinant plasmid of nucleoside triphosphate hydrolase (NTPase) gene of Toxoplasma gondii, and analyze its antigenicity. NTPase gene was amplified by PCR from RH strain of T. gondii and cloned into pGEM-T Easy vector. Positive clones were screened and identified by BglII, HindIII digestion and sequenced. The target gene was then subcloned into prokaryotic expression vector pBAD-HisB and transformed into E. coli BL21 (DE3). The expressed recombinant protein was purified with Ni-NTA agarose and further analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. NTPase-II gene was specifically amplified, and the homology of DNA sequence was 100% to that in the GenBank. SDS-PAGE showed that the recombinant NTPase protein with correct molecular weight was expressed highly in E.coli BL21 (DE3). Western blotting testified that the purified recombinant protein could be specifically recognized by mouse serum immunized with T. gondii and mouse anti-recombinant protein serum. The NTPase-II gene has been cloned and expressed in E.coli BL21 (DE3), and the purified protein of NTPase-II gene displays a specific antigenicity.
  Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases 01/2008; 25(6):447-50.
• Article: [Preparation, characterization and preliminary application of recombinant protein AP33 of trichomonas vaginalis].

  Shao-hui Liang, Hui-cong Huang, Chang-wang Pan, Wen-luan Xing, Qian Qin, Qing-yun Zhuge, Feng Tan
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  ABSTRACT: To clone ap33 gene of Trichomonas vaginalis( T. v), construct prokaryotic expression system of the gene and identify its antigenicity and immunogenicity. The total RNA was extracted from a clinical isolate Tv317 and the cDNA was synthesized by reverse transcription. The ap33 gene from cDNA of Tv317 was amplified by PCR. The nucleotide sequence of the target DNA amplification fragment was sequenced after T-A cloning. The expression vector pET32a (+) with inserted ap33 gene was constructed. Recombinant fusion protein AP33 was expressed in E. coli strain BL21DE3 induced by IPTG at different dosages. Western blotting was applied to determine immunoreactivity of the recombinant fusion protein AP33 with antibody against whole cell of T. v. Double agar diffusion was applied to determine immunogenicity of the recombinant fusion protein AP33 with rabbit antiserum immunized with the recombinant fusion protein AP33, and ELISA with antigen of T. v whole cell was applied to determine immunogenicity of the recombinant protein AP33. Positive human sera were tested by ELISA with the recombinant fusion protein AP33. High homology of nucleotide and amino acid sequences was revealed between the cloned ap33 and the corresponding gene. The recombinant protein showed a high expression level. The recombinant protein was recognized by anti-T. v polyclonal antibody from rabbit, and showed a high titer. The clinical T. v isolates showed high ap33 expression level and stimulated the production of specific antibody. Antibody against AP33 was detected in 78% of the 50 patients infected with T. v by ELISA. A prokaryotic expression system of T. v ap33 gene has been established. The expressed fusion protein AP33 shows satisfactory antigenicity and immunogenicity.
  Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases 03/2006; 24(1):31-4.
• Article: [Detection of Angiostrongylus cantonensis circulating antigen by monoclonal antibodies].

  Shao-hui Liang, Hui-cong Huang, Chang-wang Pan, Feng Tan
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  ABSTRACT: To prepare monoclonal antibodies (McAbs) against soluble antigens of adult worms of Angiostrongylus cantonensis (A. cantonensis) on the purpose to detect CAg of A. cantonensis. Female BALB/c mice were immunized with soluble antigens of adult worms of A. cantonensis and the spleen cells were fused with myeloma SP2/0 cells. The hybridoma cell strains were screened by enzyme-linked immunosorbent assay (ELISA) and Western blotting. Two McAbs (3F1 and 4H2) were applied to detect the CAg in the sera of rats and mice infected with A. cantonensis and angiostrongyliasis patients respectively by double antibody sandwich ELISA. Three McAbs against A. cantonensis adult were obtained. Two McAbs (3F1, 4H2) were identified as IgG1 and one McAb (2A2) was identified as IgM. The titers of culture fluid and ascites was 1:25,600, 1:25,600, 1:12,800 and 1:80,000, 1:80,000, 1:40,000 respectively. Western blotting results showed three McAb could be used to identify 15,000 protein of adult worms of A. cantonensis. The detection rates of the CAg in the sera of infected rats and mice were 84.2% (48/57) and 87.2% (41/47) respectively. The detection rate of the CAg in the sera of angiostrongyliasis patients was 86.4% (19/22), and no cross reactions with sera from patients with schistosomiasis, cysticercosis cellulose, paragonimiasis and trichinellosis were observed. The CAg in the sera from mice examined at different periods after infection revealed positive 2 week after inoculation and the titer of CAg peaked 4 week after inoculation. A new method of sandwich ELISA with high sensitivity and specificity to detect the serum A. cantonensis CAg has been obtained, it could be applicable to the diagnosis, observation of curative effect and epidemiology of angiostrongyliasis.
  Zhonghua yi xue za zhi 12/2005; 85(43):3057-61.
• Article: Construction of expression systems for flaA and flaB genes of Helicobacter pylori and determination of immunoreactivity and antigenicity of recombinant proteins.

  Jie Yan, Shao-Hui Liang, Ya-Fei Mao, Li-Wei Li, Shu-Ping Li
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  ABSTRACT: To clone flagellin genes A (flaA) and B (flaB) from a clinical strain of Helicobacter pylori (H pylori) and to construct prokaryotic expression systems of the genes and identify immunity of the fusion proteins. The flaA and flaB genes from a clinical H pylori isolate Y06 were amplified by high fidelity PCR. The nucleotide sequences of target DNA amplification fragments from the two genes were sequenced after T-A cloning. The recombinant expression vector pET32a inserted with flaA and flaB genes was constructed, respectively. The expressions of FlaA and FlaB fusion proteins in E. coli BL21DE3 induced by isopropylthio-beta-D-galactoside (IPTG) at different concentrations were examined by SDS-PAGE. Western blot using commercial antibodies against whole cell of H pylori and immunodiffusion assay using self-prepared rabbit antiserum against FlaA (rFlaA) or FlaB (rFlaB) recombinant proteins were applied to the determination of the fusion proteins immunity. ELISA was used to detect the antibodies against rFlaA and rFlaB in sera of 125 H pylori infected patients and to examine rFlaA and rFlaB expression in 98 clinical isolates of H pylori, respectively. In comparison with the reported corresponding sequences, the nucleotide sequence homologies of the cloned flaA and flaB genes were from 96.28-97.13% and 96.31-97.73%, and their putative amino acid sequence homologies were 99.61-99.80% and 99.41-100% for the two genes, respectively. The output of rFlaA and rFlaB expressed by pET32a-flaA-BL21DE3 and pET32a-flaB-BL21DE3 systems was as high as 40-50% of the total bacterial proteins. Both rFlaA and rFlaB were able to combine with the commercial antibodies against whole cell of H pylori and to induce rabbits to produce specific antibodies with the same 1:2 immunodiffusion titers after the animals were immunized with the two recombinant proteins. Ninety-eight and zero point 4 and 92.80% of the serum samples from 125 patients infected with H pylori were positive for rFlaA and rFlaB antibodies, respectively. One hundred percent and 98.98% of the 98 tested isolates of H pylori were detectable for rFlaA and rFlaB epitopes, respectively. Two prokaryotic expression systems with high efficiency of H pylori flaA and flaB genes were successfully established. The expressed rFlaA and rFlaB showed satisfactory immunoreactivity and antigenicity. High frequencies of FlaA and FlaB expression in different H pylori clinical strains and the general existence of specific antibodies against FlaA and FlaB in H pylori infected patients strongly indicate that FlaA and FlaB are excellent antigen candidates for developing H pylori vaccine.
  World Journal of Gastroenterology 11/2003; 9(10):2240-50. · 2.47 Impact Factor
• Article: [Cloning, expression and identification of flaB gene from a clinical isolate of Helicobacter pylori].

  Shao-Hui Liang, Ya-Fei Mao, Jie Yan
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  ABSTRACT: To clone Helicobacter pylori flagellin B gene (flaB) to construct prokaryotic expression system of the gene and to identify immunity of the fusion protein. The flaB gene from a clinical isolate Y06 of H.pylori was amplified by high fidelity PCR. The nucleotide sequence of the target DNA amplification fragment was sequenced after T-A cloning. The expression vector pET32a with inserted flaB gene was constructed. FlaB fusion protein was expressed in E.coli strain BL21DE3 inducted by IPTG at different dosages. Western blot was applied to determine immunoreactivity and immunogenicity of the fusion protein and antibody against whole cell of H.pylori and rabbit antiserum immunized with the fusion protein, respectively. In comparison with the reported corresponding sequences, the homology of nucleotide sequence of the cloned flaB gene was from 96.31% approximate, equals 97.73%, while the homology of its putative amino acid sequence was as high as 99.41% approximate, equals 100%. The expression output of FlaB fusion protein in pET32a-flaB-BL21DE3 system was approximately 40% of the total bacterial proteins. FlaB fusion protein was able to combine with antibody against whole cell of H.pylori and induce rabbit to produce specific antibody with high titer after the animal was immunized with the protein. A prokaryotic expression system of H. pylori flaB gene with high efficiency has been established successfully. The expressed FlaB fusion protein with satisfactory immunogenicity and immunoreactivity can be used as antigen in H.pylori vaccine and detect kit.
  Zhejiang da xue xue bao. Yi xue ban = Journal of Zhejiang University. Medical sciences 03/2003; 32(1):13-6.