Objective To detect the specific mutations in rpoB gene of Mycobacterium tuberculosis by oligonucleotide microarray. Methods Four wild-type and 8 mutant probes were used to detect rifampin resistant strains. Target DN...Objective To detect the specific mutations in rpoB gene of Mycobacterium tuberculosis by oligonucleotide microarray. Methods Four wild-type and 8 mutant probes were used to detect rifampin resistant strains. Target DNA of M. tuberculosis was amplified by PCR, hybridized and scanned. Direct sequencing was performed to verify the results of oligonucleotide microarray Results Of the 102 rifampin-resistant strains 98 （96.1%） had mutations in the rpoB genes. Conclusion Oligonucleotide microarray with mutation-specific probes is a reliable and useful tool for the rapid and accurate diagnosis of rifampin resistance in M. tuberculosis isolates.展开更多
Objectives To evaluate the immunogenicity of Mycobacterium intracellulare proteins and determine the cross-reactive proteins between M.intracellulare and M.tuberculosis.Methods Protein extracts from M.intracellulare w...Objectives To evaluate the immunogenicity of Mycobacterium intracellulare proteins and determine the cross-reactive proteins between M.intracellulare and M.tuberculosis.Methods Protein extracts from M.intracellulare were used to immunize BALB/c mice.The antigens were evaluated using cellular and humoral immunoassays.The common genes between M.intracellular and M.tuberculosis were identified using genome-wide comparative analysis,and cross-reactive proteins were screened using immunoproteome microarrays.Results Immunization with M.intracellulare proteins induced significantly higher levels of the cytokines interferon-γ(IFN-γ),interleukin-2(IL-2),interleukin-12(IL-12),interleukin-6(IL-6)and immunoglobulins IgG,IgG1,IgM,and IgG2a in mouse serum.Bone marrow-derived macrophages isolated from mice immunized with M.intracellulare antigens displayed significantly lower bacillary loads than those isolated from mice immunized with adjuvants.Whole-genome sequence analysis revealed 396 common genes between M.intracellulare and M.tuberculosis.Microchip hybridization with M.tuberculosis proteins revealed the presence of 478 proteins in the serum of mice immunized with M.intracellulare protein extracts.Sixty common antigens were found using both microchip and genomic comparative analyses.Conclusion This is the advanced study to investigate the immunogenicity of M.intracellulare proteins and the cross-reactive proteins between M.intracellulare and M.tuberculosis.The results revealed the presence of a number of cross-reactive proteins between M.intracellulare and M.tuberculosis.Therefore,this study provides a new way of identifying immunogenic proteins for use in tuberculosis vaccines against both M.intracellulare and M.tuberculosis in future.展开更多
目的探讨m1A RNA甲基化相关基因和血浆m1A甲基化水平对结肠腺癌(colorectal adenocarcinoma,COAD)的诊断效能,为COAD早期诊断提供新的方案。方法通过UALCAN、The Human Protein Atlas和TCGA-GTEx数据库,分析COAD组织和正常结肠组织中m1...目的探讨m1A RNA甲基化相关基因和血浆m1A甲基化水平对结肠腺癌(colorectal adenocarcinoma,COAD)的诊断效能,为COAD早期诊断提供新的方案。方法通过UALCAN、The Human Protein Atlas和TCGA-GTEx数据库,分析COAD组织和正常结肠组织中m1A相关基因mRNA和蛋白水平的差异表达。利用ELISA法检测收集于我院初诊的COAD患者和正常人血浆中m1A甲基化水平。结合COAD临床病理特征分析m1A甲基化对COAD的诊断效能。结果m1A编码器和读码器基因的蛋白水平和mRNA水平在COAD组织中的表达显著上调,其中以TRMT6和TRMT10C两个编码器表达升高最为显著。两个编码器基因均可作为COAD诊断,尤其是早期诊断标志物,且其AUC均达到0.9以上。m1A总体甲基化水平在COAD血浆中明显升高,并可作为早期COAD的诊断标志物。结论m1A编码器基因和血浆m1A在COAD中明显升高,有望成为一种新的早期COAD诊断标志物。展开更多
基金supported by the grant from the National Natural Science Foundation of China (No. 30400018)
文摘Objective To detect the specific mutations in rpoB gene of Mycobacterium tuberculosis by oligonucleotide microarray. Methods Four wild-type and 8 mutant probes were used to detect rifampin resistant strains. Target DNA of M. tuberculosis was amplified by PCR, hybridized and scanned. Direct sequencing was performed to verify the results of oligonucleotide microarray Results Of the 102 rifampin-resistant strains 98 （96.1%） had mutations in the rpoB genes. Conclusion Oligonucleotide microarray with mutation-specific probes is a reliable and useful tool for the rapid and accurate diagnosis of rifampin resistance in M. tuberculosis isolates.
基金supported by National Science and Technology Major Project of China[2018ZX10731301-002]。
文摘Objectives To evaluate the immunogenicity of Mycobacterium intracellulare proteins and determine the cross-reactive proteins between M.intracellulare and M.tuberculosis.Methods Protein extracts from M.intracellulare were used to immunize BALB/c mice.The antigens were evaluated using cellular and humoral immunoassays.The common genes between M.intracellular and M.tuberculosis were identified using genome-wide comparative analysis,and cross-reactive proteins were screened using immunoproteome microarrays.Results Immunization with M.intracellulare proteins induced significantly higher levels of the cytokines interferon-γ(IFN-γ),interleukin-2(IL-2),interleukin-12(IL-12),interleukin-6(IL-6)and immunoglobulins IgG,IgG1,IgM,and IgG2a in mouse serum.Bone marrow-derived macrophages isolated from mice immunized with M.intracellulare antigens displayed significantly lower bacillary loads than those isolated from mice immunized with adjuvants.Whole-genome sequence analysis revealed 396 common genes between M.intracellulare and M.tuberculosis.Microchip hybridization with M.tuberculosis proteins revealed the presence of 478 proteins in the serum of mice immunized with M.intracellulare protein extracts.Sixty common antigens were found using both microchip and genomic comparative analyses.Conclusion This is the advanced study to investigate the immunogenicity of M.intracellulare proteins and the cross-reactive proteins between M.intracellulare and M.tuberculosis.The results revealed the presence of a number of cross-reactive proteins between M.intracellulare and M.tuberculosis.Therefore,this study provides a new way of identifying immunogenic proteins for use in tuberculosis vaccines against both M.intracellulare and M.tuberculosis in future.
文摘目的探讨m1A RNA甲基化相关基因和血浆m1A甲基化水平对结肠腺癌(colorectal adenocarcinoma,COAD)的诊断效能,为COAD早期诊断提供新的方案。方法通过UALCAN、The Human Protein Atlas和TCGA-GTEx数据库,分析COAD组织和正常结肠组织中m1A相关基因mRNA和蛋白水平的差异表达。利用ELISA法检测收集于我院初诊的COAD患者和正常人血浆中m1A甲基化水平。结合COAD临床病理特征分析m1A甲基化对COAD的诊断效能。结果m1A编码器和读码器基因的蛋白水平和mRNA水平在COAD组织中的表达显著上调,其中以TRMT6和TRMT10C两个编码器表达升高最为显著。两个编码器基因均可作为COAD诊断,尤其是早期诊断标志物,且其AUC均达到0.9以上。m1A总体甲基化水平在COAD血浆中明显升高,并可作为早期COAD的诊断标志物。结论m1A编码器基因和血浆m1A在COAD中明显升高,有望成为一种新的早期COAD诊断标志物。
