AIM： To detect the common intestinal pathogenic bacteria quickly and accurately.METHODS： A rapid （〈3 h） experimental procedure was set up based upon the gene chip technology, Target genes were amplified and hybri...AIM： To detect the common intestinal pathogenic bacteria quickly and accurately.METHODS： A rapid （〈3 h） experimental procedure was set up based upon the gene chip technology, Target genes were amplified and hybridized by oligonucleotide microarrays.RESULTS： One hundred and seventy strains of bacteria in pure culture belonging to 11 genera were successfully discriminated under comparatively same conditions, and a series of specific hybridization maps corresponding to each kind of bacteria were obtained. When this method was applied to 26 divided cultures, 25 （96.2%） were identified.CONCLUSION： Salmonella sp., Escherichia coli, Shigella sp., Listeria monocytogenes, Vibrio parahaemolyticus, Staphylococcus aureus , Proteus sp., Bacillus cereus, Vibrio cholerae, Enterococcus faecalis, Yersinia enterocolitica, and Campylobacter jejuni can be detected and identified by our microarrays. The accuracy, range, and discrimination power of this assay can be continually improved by adding further oligonudeotides to the arrays without any significant increase of complexity or cost.展开更多
AIM： TO determine the genotype distribution of hepatitis B virus （HBV） with a newly oligonucleotide chip assay among the HBV carriers in Eastern China. METHODS： An assay using oligonucleotide chip was developed fo...AIM： TO determine the genotype distribution of hepatitis B virus （HBV） with a newly oligonucleotide chip assay among the HBV carriers in Eastern China. METHODS： An assay using oligonucleotide chip was developed for detection of HBV genotypes in serum samples from HBV DNA-positive patients in Eastern China. This method is based on the principle of reverse hybridization with Cy5-labeled amplicons hybridizing to type-specific oligonucleotide probes that are immobilized on slides. The results of 80 randomly chosen sera were confirmed by direct sequencing. RESULTS： HBV genotype B, C and mixed genotype were detected in 400 serum samples, accounting for 8.3% （n = 33）, 83.2% （n = 333）, and 8.5% （n = 34）, respectively. The evaluation of the oligonucleotide assay showed 100% concordance with the amplicon phylogenetic analysis except 9 mixed genotype infections undetected by sequencing. CONCLUSION： The study indicates that HBV genotype C and B prevail in the Eastern China. It is suggested that the oligonucleotide chip is a reliable and convenient tool for the detection of HBV genotyping.展开更多
目的:建立乙型肝炎病毒(hepatitis B virus,HBV)B基因型亚型的检测方法,并对其进行临床研究.方法:根据GenBank中20株Ba亚型,10株B j亚型以及25株C型全长基因组序列设计Ba、Bj亚型特异探针以及巢式PCR引物.采用反向杂交技术将B亚型特异...目的:建立乙型肝炎病毒(hepatitis B virus,HBV)B基因型亚型的检测方法,并对其进行临床研究.方法:根据GenBank中20株Ba亚型,10株B j亚型以及25株C型全长基因组序列设计Ba、Bj亚型特异探针以及巢式PCR引物.采用反向杂交技术将B亚型特异探针固定在芯片上,通过与地高辛标记的扩增产物杂交检测B型HBV亚型.对镇江地区200例B基因型HBV血清样本进行亚型分析并对患者HBV DNA YMDD变异进行检测,通过对部分血清中的HBV DNA进行序列分析验证该方法的准确性.结果:200例B基因型HBV患者中,Ba亚型184例(92%),Bj亚型16例(8%).在持续使用拉米呋啶的80例患者中,Bj亚型6(0%)例,均无YMDD变异,Ba亚型74例,YMDD变异21(28.4%)例.结论:镇江地区B基因型HBV以Ba亚型为主,Ba、Bj亚型在服用拉米呋啶1年后发生YMDD变异的几率并无统计学上的显著差异.展开更多
Based on the c DNA sequences from hyper variable(HV) regions of identified 52 S-alleles in Oriental pear cultivars, S-RNase c DNA probes were designed, and a c DNA microarray for S-RNase detections was established. Ea...Based on the c DNA sequences from hyper variable(HV) regions of identified 52 S-alleles in Oriental pear cultivars, S-RNase c DNA probes were designed, and a c DNA microarray for S-RNase detections was established. Each microarray contained 240 sites from 55 c DNA probes, including all specific c DNA sequences from the HV regions of the S-alleles. Using the c DNA of pistils of tested pear cultivars as template and Cy3 fluorescently labeling primers by PCR amplification, microarray hybridization detected the S-genotype of each pear cultivar. The genotypes inferred from the c DNA microarray hybridization signals of pear cultivars such as ‘Lijiang Huangsuanli', ‘Xiuyu', ‘Midu Yuli', ‘Baimianli', and ‘Deshengxiang' were similar to the known genotypes of all tested cultivars. The S-RNase c DNA microarrays and the oligonucleotide gene chips were then used to conduct parallel testing of 24 P. pyrifolia cultivars with unknown S-genotypes. In conclusion, the construction of c DNA microarrays has further improved the pear S-RNase detection platform.展开更多
目的设计、制作一种微型化寡核苷酸阵列芯片,评价快速鉴定肠出血性大肠杆菌O157:H7的效果。方法多重PCR扩增大肠杆菌O157:H7七个特异性基因位点(rfbEf、licH7i、ntimin、Shiga-like toxins I and II、hemolysin A和uidA),通过PCR反应掺...目的设计、制作一种微型化寡核苷酸阵列芯片,评价快速鉴定肠出血性大肠杆菌O157:H7的效果。方法多重PCR扩增大肠杆菌O157:H7七个特异性基因位点(rfbEf、licH7i、ntimin、Shiga-like toxins I and II、hemolysin A和uidA),通过PCR反应掺入SpectrumOrangTM-dUTP获取荧光标记的靶序列,与制备的芯片寡核苷酸探针杂交。结果寡核苷酸阵列芯片检测结果与试验预期相符,获取的杂交图分辨效果明显优于多重PCR琼脂糖凝胶电泳。结论基于玻片的寡核苷酸阵列芯片制作简便,鉴定病原菌检测细菌毒力因子快速、灵敏、特异,有良好的应用前景。展开更多
基金Supported by the National High Technology ResearchDevelopment Program of China (863 Program), No.2002AA2Z2011
文摘AIM： To detect the common intestinal pathogenic bacteria quickly and accurately.METHODS： A rapid （〈3 h） experimental procedure was set up based upon the gene chip technology, Target genes were amplified and hybridized by oligonucleotide microarrays.RESULTS： One hundred and seventy strains of bacteria in pure culture belonging to 11 genera were successfully discriminated under comparatively same conditions, and a series of specific hybridization maps corresponding to each kind of bacteria were obtained. When this method was applied to 26 divided cultures, 25 （96.2%） were identified.CONCLUSION： Salmonella sp., Escherichia coli, Shigella sp., Listeria monocytogenes, Vibrio parahaemolyticus, Staphylococcus aureus , Proteus sp., Bacillus cereus, Vibrio cholerae, Enterococcus faecalis, Yersinia enterocolitica, and Campylobacter jejuni can be detected and identified by our microarrays. The accuracy, range, and discrimination power of this assay can be continually improved by adding further oligonudeotides to the arrays without any significant increase of complexity or cost.
文摘AIM： TO determine the genotype distribution of hepatitis B virus （HBV） with a newly oligonucleotide chip assay among the HBV carriers in Eastern China. METHODS： An assay using oligonucleotide chip was developed for detection of HBV genotypes in serum samples from HBV DNA-positive patients in Eastern China. This method is based on the principle of reverse hybridization with Cy5-labeled amplicons hybridizing to type-specific oligonucleotide probes that are immobilized on slides. The results of 80 randomly chosen sera were confirmed by direct sequencing. RESULTS： HBV genotype B, C and mixed genotype were detected in 400 serum samples, accounting for 8.3% （n = 33）, 83.2% （n = 333）, and 8.5% （n = 34）, respectively. The evaluation of the oligonucleotide assay showed 100% concordance with the amplicon phylogenetic analysis except 9 mixed genotype infections undetected by sequencing. CONCLUSION： The study indicates that HBV genotype C and B prevail in the Eastern China. It is suggested that the oligonucleotide chip is a reliable and convenient tool for the detection of HBV genotyping.
文摘目的:建立乙型肝炎病毒(hepatitis B virus,HBV)B基因型亚型的检测方法,并对其进行临床研究.方法:根据GenBank中20株Ba亚型,10株B j亚型以及25株C型全长基因组序列设计Ba、Bj亚型特异探针以及巢式PCR引物.采用反向杂交技术将B亚型特异探针固定在芯片上,通过与地高辛标记的扩增产物杂交检测B型HBV亚型.对镇江地区200例B基因型HBV血清样本进行亚型分析并对患者HBV DNA YMDD变异进行检测,通过对部分血清中的HBV DNA进行序列分析验证该方法的准确性.结果:200例B基因型HBV患者中,Ba亚型184例(92%),Bj亚型16例(8%).在持续使用拉米呋啶的80例患者中,Bj亚型6(0%)例,均无YMDD变异,Ba亚型74例,YMDD变异21(28.4%)例.结论:镇江地区B基因型HBV以Ba亚型为主,Ba、Bj亚型在服用拉米呋啶1年后发生YMDD变异的几率并无统计学上的显著差异.
基金supported by the National Natural Science Foundation of China(31272124)
文摘Based on the c DNA sequences from hyper variable(HV) regions of identified 52 S-alleles in Oriental pear cultivars, S-RNase c DNA probes were designed, and a c DNA microarray for S-RNase detections was established. Each microarray contained 240 sites from 55 c DNA probes, including all specific c DNA sequences from the HV regions of the S-alleles. Using the c DNA of pistils of tested pear cultivars as template and Cy3 fluorescently labeling primers by PCR amplification, microarray hybridization detected the S-genotype of each pear cultivar. The genotypes inferred from the c DNA microarray hybridization signals of pear cultivars such as ‘Lijiang Huangsuanli', ‘Xiuyu', ‘Midu Yuli', ‘Baimianli', and ‘Deshengxiang' were similar to the known genotypes of all tested cultivars. The S-RNase c DNA microarrays and the oligonucleotide gene chips were then used to conduct parallel testing of 24 P. pyrifolia cultivars with unknown S-genotypes. In conclusion, the construction of c DNA microarrays has further improved the pear S-RNase detection platform.
文摘目的设计、制作一种微型化寡核苷酸阵列芯片,评价快速鉴定肠出血性大肠杆菌O157:H7的效果。方法多重PCR扩增大肠杆菌O157:H7七个特异性基因位点(rfbEf、licH7i、ntimin、Shiga-like toxins I and II、hemolysin A和uidA),通过PCR反应掺入SpectrumOrangTM-dUTP获取荧光标记的靶序列,与制备的芯片寡核苷酸探针杂交。结果寡核苷酸阵列芯片检测结果与试验预期相符,获取的杂交图分辨效果明显优于多重PCR琼脂糖凝胶电泳。结论基于玻片的寡核苷酸阵列芯片制作简便,鉴定病原菌检测细菌毒力因子快速、灵敏、特异,有良好的应用前景。
