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Novel biosensor-based microarray assay for detecting rs8099917 and rs12979860 genotypes

World Journal of Gastroenterology 2012年第44期18卷 6481-6488页

作者： Pei-Yuan Li Xiao-Jun Zhou Lan Yao Xin-Hua Fang Jiang-Nan Ren Jia-Wu Song Department of Gastroenterology the Fifth Affili- ated Hospital of Stm Yat-Sen University Zhuhai 519000 Guang- dong Province China Department of Emergency the Fifth Affiliated Hospital of Sun Yat-Sen University Zhuhai 519000 Guangdong Province China

AIM:To evaluate a novel biosensor-based microarray(BBM) assay for detecting rs12979860 and rs8099917 ***:Four probes specific for rs8099917c/T or rs12979860G/T detection and three sets of quality control probes were designed,constructed and arrayed on an optical biosensor to develop a microarray *** sets of primers were used in a one tube polymerase chain reaction(PcR) system to amplify two target fragments *** biosensor microarray contained probes that had been sequenced to confirm that they included the rs8099917c/T or rs12979860G/T alleles of interest and could serve as the specific assay *** addition to rehybridization of four probes of known sequence,a total of 40 clinical samples collected from hepatitis c seropositive patients were also *** target fragments of all 40 samples were amplified in a 50 μL PcR *** μL of each amplicon was tested by BBM assay,and another 40 μL was used for *** agreement of the results obtained by the two methods was tested statistically using the kappa *** sensitivity of the BBM assay was evaluated using serial dilutions of ten clinical blood samples containing 10 3-10 4 white cells/μ***:As shown by polyacrylamide gel electrophoresis,two target segments of the interleukin 28Bassociated polymorphisms(SNPs) were successfully amplified in the one-tube PcR *** lengths of the two amplified fragments were consistent with the known length of the target sequences,137 and 159 *** hybridization of the PcR amplicons with the probes located on the BBM array,the signals of each allele of both the rs8099917 SNPs and rs12979860 SNPs were observed simultaneously and were clearly visible by the unaided *** signals were distinct from each other,could be interpreted visually,and accurately recorded using an ordinary digital *** evaluate the specificity of the assay,both the plasmids and clinical samples were applied to the ***,30 P

关键词： Biosensor-based microarray hepatitis c vi-rus rs8099917 rs12979860 Detection Assay

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constant serum levels of secreted asialoglycoprotein receptor sH2a and decrease with cirrhosis

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World Journal of Gastroenterology 2011年第48期17卷 5305-5309页

作者： Ron Benyair Maria Kondratyev Elena Veselkin Sandra Tol-chinsky Marina Shenkman Yoav Lurie Gerardo Z Lederkremer Department of cell Research and Immunology George Wise Faculty of Life Sciences Tel Aviv University Liver Unit Department of Gastroenterology Tel Aviv Sourasky Medical Center

AIM: To investigate the existence and levels of sH2a, a soluble secreted form of the asialoglycoprotein receptor in human serum. METHODS: Production of recombinant sH2a and development of a monoclonal antibody and an enzyme-linked immunosorbent assay (ELISA). This assay was used to determine the presence and concentration of sH2a in human sera of individuals of both sexes and a wide range of ages. RESULTS: The recombinant protein was produced successfully and a specific ELISA assay was developed. The levels of sH2a in sera from 62 healthy individuals variedminimally (147 ± 19 ng/mL). In contrast, 5 hepatitis c patients with cirrhosis showed much decreased sH2a levels (50 ± 9 ng/mL). cONcLUSION: constant sH2a levels suggest constitutive secretion from hepatocytes in healthy individuals. This constant level and the decrease with cirrhosis suggest a diagnostic potential.

关键词： Asialoglycoprotein receptor hepatitis c vi-rus Liver function cirrhosis Liver diagnosis

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