Rapid and high throughput detection of HBV YMDD mutants with fluorescence polarization
Rapid and high throughput detection of HBV YMDD mutants with fluorescence polarization作者机构：Institute of Genetic Diagnosis Fourth Military Medical University Xi'an 710032 Shannxi Province China. yujiebai@***
出 版 物：《World Journal of Gastroenterology》 (世界胃肠病学杂志（英文版）)
年 卷 期：2003年第9卷第10期
页 面：2344-2347页
核心收录：
学科分类：1004[医学-公共卫生与预防医学(可授医学、理学学位)] 1002[医学-临床医学] 1010[医学-医学技术（可授医学、理学学位）] 100401[医学-流行病与卫生统计学] 100215[医学-康复医学与理疗学] 10[医学]
基 金：Shen Zhen Syno Gene Digital Co.Ltd
主 题：TDI-FP YMDD 乙型肝炎病毒 基因变异 血清标本
摘 要：AIM: To develop a simple and rapid detection of HBV gene variants and prediction of lamivudine-resistance in ***: Initially, plasmids harboring the wild-type or mutant HBV DNA fragments were used in a model *** technique was then applied to clinical samples for an analysis of YMDD mutations. The sera were extracted from chronic hepatitis patients who had received lamivudine treatment for more than one year. P region gene of HBV was amplified by polymerase chain reaction. The excess primers and dNTPs in PCR products were removed by cleaning-up reagents. Template-directed dye-terminator incorporation reaction was performed and R110 or TAMRA labeled acyclo-terminator was added on the 3 end of TDIprimer specifically. Fluorescence polarization value was measured with Victor 2 multilabel counter and the genotypes of HBV were ***: The YMDD genotypes in recombined positive plasmid and 56 serum samples of HBV infected patients were analyzed by using our TDI-FP method and the specificity and sensitivity were confirmed by DNA sequencing. Five of 56 serum samples showed YVDD phenotype (9%), including 1 YMDD and YVDD mixed infection. Four of 56 showed YIDD phenotype (7.1%).CONCLUSION: This is a simple, rapid, low cost and high throughput assay to detect HBV polymerase gene variants and suitable for large-scale screening and prediction of the lamivudine-resistance in clinical samples.
维普期刊数据库