Objective To establish and evaluate a real-time PCR assay to detect Mycoplasma pneumoniae （M.pneumoniae） in clinical specimens. Methods By analysing the whole pl gene sequence of 60 M.pneurnoniae clinical isolates i...Objective To establish and evaluate a real-time PCR assay to detect Mycoplasma pneumoniae （M.pneumoniae） in clinical specimens. Methods By analysing the whole pl gene sequence of 60 M.pneurnoniae clinical isolates in Beijing of China, an optimized real-time PCR assay （MpP1） using pl gene conserved region was designed. The specificity and sensitivity of this assay were evaluated and compared with other two reported assays （RepMpl and Mp181） using 40 positive and 100 negative clinical specimens. Results The detection limit of the new assay was 8.1 fg （about 1-3CFU） M.pneumoniae DNA. The sensitivity of MpP1, RepMpl, and Mp181 assays appeared to be 100%, 100%, and 85%, respectively. Conclusion MpP1 assay is suitable for the detection of M.pneumoniae in Chinese clinical specimens.展开更多
Objective Secreted proteins of Helicobacter pylori(H.pylori)interact with gastric epithelium cells and may contribute to cell damage.Considering the fact that HPO17S and hypothetical conserved protein HP1286 are inclu...Objective Secreted proteins of Helicobacter pylori(H.pylori)interact with gastric epithelium cells and may contribute to cell damage.Considering the fact that HPO17S and hypothetical conserved protein HP1286 are included in the group and that HP0175 is a well-known apoptosis-induced factor,the present study aims to clarify whether HP1286 plays a role in bacterial pathogenicity or even functions as an apoptosis-induced factor in human stomach.Methods Two genes encoding HP1286 and HPO175 were cloned into pET32a vector and expressed as recombinant proteins in Escherichia coil(E.coil)BL21.Signal peptide sequences were not included.The recombinant proteins were purified with His SpinTrap and desalted by using HiTrap Desalting.Immunoreactivity of the proteins was determined by Western blot.Human gastric epithelial cell AGS was challenged with these endotoxin-free proteins;and apoptosis of cells was assayed with the Cell Death ELISA kit.Results Recombinant proteins and their respective products whose N-terminal his-tag were removed with thrombin were recognized by serum from the patient infected with H.pylori.Apoptotic AGS cells challenged by HP1286 of H.pylori 26695 were four times more than untreated cells.In addition,apoptosis-induced ability of HP1286 of SS1 was not as strong as that of H.pylori 26695 strain.Conclusion HP1286 of H.pylori 26695 induces apoptosis of AGS cells in a time-dependent manner,however the apoptosis-induced ability of HP1286 may differ due to variations between different strains.展开更多
Objective To investigate the protein expression profiles of the major food‐borne pathogen Campylobacter jejuni NCTC11168.Methods Membrane and soluble cellular proteins were extracted from the genome‐sequenced C.jeju...Objective To investigate the protein expression profiles of the major food‐borne pathogen Campylobacter jejuni NCTC11168.Methods Membrane and soluble cellular proteins were extracted from the genome‐sequenced C.jejuni strain NCTC11168.Protein expression profiles were determined using two‐dimensional gel electrophoresis（2‐DE）.All the detected spots on the 2‐DE map were subjected to matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry（MALDI‐TOF/TOF） analysis.Results A total of 537 and 333 spots were detected from the whole cell and membrane‐associated proteins of C.jejuni NCTC11168 cultured on Columbia agar medium at 42 ℃ by 2‐DE and Coomassie Brilliant Blue staining,respectively.Analyses of whole cell and membrane‐associated proteins included 399 and 133 spots,respectively,which included 182 and 53 functional proteins identified by MALDI‐TOF/TOF analysis.Conclusion The comprehensive expression protein profiles of C.jeuni NCTC11168 obtained in this study will be useful for elucidating the roles of these proteins in further pathogenesis investigation.展开更多
This study aimed to determine the in vitro activity of quinupristin-alfopristin against Streptococcus sp. isolated in China. This agent is not yet available for clinical use, but it has been tested against a high prop...This study aimed to determine the in vitro activity of quinupristin-alfopristin against Streptococcus sp. isolated in China. This agent is not yet available for clinical use, but it has been tested against a high proportion of resistant Staphylococcus oureus strains. A total of 156 streptococcal isolates, which were recovered from various geographic areas and diseases, were tested using the Etest （AB Biodisk, Solna, Sweden）. Quinupristin-alfopristin showed excellent activity against all of the tested streptococci isolates. These results provide useful data for the clinical use of quinupristin-alfopristin in China.展开更多
基金supported by the National Key Program for Infectious Diseases of China,No.2008ZX10004-002
文摘Objective To establish and evaluate a real-time PCR assay to detect Mycoplasma pneumoniae （M.pneumoniae） in clinical specimens. Methods By analysing the whole pl gene sequence of 60 M.pneurnoniae clinical isolates in Beijing of China, an optimized real-time PCR assay （MpP1） using pl gene conserved region was designed. The specificity and sensitivity of this assay were evaluated and compared with other two reported assays （RepMpl and Mp181） using 40 positive and 100 negative clinical specimens. Results The detection limit of the new assay was 8.1 fg （about 1-3CFU） M.pneumoniae DNA. The sensitivity of MpP1, RepMpl, and Mp181 assays appeared to be 100%, 100%, and 85%, respectively. Conclusion MpP1 assay is suitable for the detection of M.pneumoniae in Chinese clinical specimens.
基金supported by the Major Technology Project as part of"Prevention and Control of Major Infectious Diseases including AIDS and Viral Hepatitis"(No.2008ZX10004-002)
文摘Objective Secreted proteins of Helicobacter pylori(H.pylori)interact with gastric epithelium cells and may contribute to cell damage.Considering the fact that HPO17S and hypothetical conserved protein HP1286 are included in the group and that HP0175 is a well-known apoptosis-induced factor,the present study aims to clarify whether HP1286 plays a role in bacterial pathogenicity or even functions as an apoptosis-induced factor in human stomach.Methods Two genes encoding HP1286 and HPO175 were cloned into pET32a vector and expressed as recombinant proteins in Escherichia coil(E.coil)BL21.Signal peptide sequences were not included.The recombinant proteins were purified with His SpinTrap and desalted by using HiTrap Desalting.Immunoreactivity of the proteins was determined by Western blot.Human gastric epithelial cell AGS was challenged with these endotoxin-free proteins;and apoptosis of cells was assayed with the Cell Death ELISA kit.Results Recombinant proteins and their respective products whose N-terminal his-tag were removed with thrombin were recognized by serum from the patient infected with H.pylori.Apoptotic AGS cells challenged by HP1286 of H.pylori 26695 were four times more than untreated cells.In addition,apoptosis-induced ability of HP1286 of SS1 was not as strong as that of H.pylori 26695 strain.Conclusion HP1286 of H.pylori 26695 induces apoptosis of AGS cells in a time-dependent manner,however the apoptosis-induced ability of HP1286 may differ due to variations between different strains.
基金supported by The General Program of National Natural Science Foundation of China(81071314)
文摘Objective To investigate the protein expression profiles of the major food‐borne pathogen Campylobacter jejuni NCTC11168.Methods Membrane and soluble cellular proteins were extracted from the genome‐sequenced C.jejuni strain NCTC11168.Protein expression profiles were determined using two‐dimensional gel electrophoresis（2‐DE）.All the detected spots on the 2‐DE map were subjected to matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry（MALDI‐TOF/TOF） analysis.Results A total of 537 and 333 spots were detected from the whole cell and membrane‐associated proteins of C.jejuni NCTC11168 cultured on Columbia agar medium at 42 ℃ by 2‐DE and Coomassie Brilliant Blue staining,respectively.Analyses of whole cell and membrane‐associated proteins included 399 and 133 spots,respectively,which included 182 and 53 functional proteins identified by MALDI‐TOF/TOF analysis.Conclusion The comprehensive expression protein profiles of C.jeuni NCTC11168 obtained in this study will be useful for elucidating the roles of these proteins in further pathogenesis investigation.
基金supported by funding from the China Mega-Project for Infectious Disease(2011ZX10004-001)a grant from the National Technology R&D Program in the 12th Five-Year Plan of China(2012BAI06B02)
文摘This study aimed to determine the in vitro activity of quinupristin-alfopristin against Streptococcus sp. isolated in China. This agent is not yet available for clinical use, but it has been tested against a high proportion of resistant Staphylococcus oureus strains. A total of 156 streptococcal isolates, which were recovered from various geographic areas and diseases, were tested using the Etest （AB Biodisk, Solna, Sweden）. Quinupristin-alfopristin showed excellent activity against all of the tested streptococci isolates. These results provide useful data for the clinical use of quinupristin-alfopristin in China.
