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
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
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.
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.
