一种国产基因芯片用于乙型肝炎病毒P基因突变的检测

目的：验证国产基因芯片检测HBV—P基因突变的准确性和敏感性。方法：选择14例服用拉米夫定48周时HBV—DNA定量检测仍阳性的慢性乙型肝炎病人的血清标本，用HBV基因突变检测基因芯片检测患者血清中HBV—DNA聚合酶基因，观察发生YMDD变异的情况。同时用聚合酶链反应(PCR）技术扩增上述血清标本中编码HBV—DNA聚合酶的P基因片段并直接测序，以对比检查P基因发生YMDD变异的情况。结果：14例患者血清标本用基因芯片均检出HBV—DNA，9例为野生株，5例发生YMDD变异，其中3例为M552I(YIDD)变异，2例为M552V(YVDD)变异，同时均伴有L528M变异。与用PCR扩增产物直接测序的结果完全相同。结论：国产HBV基因突变检测芯片具有特异性强、灵敏性高、快速、简便等优点，可以替代繁琐的DNA序列测定和分析，可作为临床监测HBV—DNA变异的常规方法。

Detection on gene mutation of HBV with microarray genechip

Objective: To verify the quality of the mciroarr ay genechips(made in China) that can be used to detect the gene mutation of HBV. Methods: 14 cases of chronic hepatitis B were selected. Aft er they had taken lamivudine for 48 weeks, the HBV - DNA in their blood were st ill positive based on quantitative analysis. Then their blood samples were colle cted and detected for YMDD motif mutation in HBV - DNA polymerase gene by the m icroarray genechips for HBV mutation. At the same time, P gene of HBV in these b lood samples were amplified by PCR, and the gene sequences were detected to inve stigate the YMDD motif mutation. Results: HBV - DNA was fou nd in the blood samples from 14 cases detected by microarray genechips. Among th em, 9 cases were wild type, YMDD mutation were found in the other 5 ones. Among these 5 cases, 3 ones were M552I(YIDD) mutation and the other 2 ones were M552V( YVDD) mutation accompanied by L528M mutation. These results were the same as tho se obtained by sequencing of the PCR products. Conclusion: T he microarray gengchip for HBV gene mutation detection made in China was specifi c, sensitive and convenient, and it can be used to replace DNA sequencing and ma ybe become a routine method to detect HBV - DNA mutation.