State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai 200237, PR China; Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Science, 867 Changning Road, Shanghai 200050, PR China.
Hepatol Res. 2006 Mar;34(3):150-5. doi: 10.1016/j.hepres.2005.12.007. Epub 2006 Feb 24.
A novel method based on ligase detection reaction (LDR) coupled with polymerase chain reaction (PCR) was used in this study to detect low abundant YIDD mutants in hepatitis B virus (HBV), which was able to detect 10(2) mutants from 10(8) wild type copies in plasmids and 10(3) from 10(7) in clinical serum samples. Direct sequencing and microarray hybridization were selected for sensitivity comparison together with this assay. By direct sequencing, 6 of 50 clinical specimens infected with HBV were found to be YIDD and the others to be YMDD, while by microarray hybridization, 28 specimens were detected to contain both YMDD and YIDD, and the others contained YMDD only, and with this assay, 31 specimens containing both YMDD and YIDD were detected with the others containing YMDD only, which indicated that the assay was relatively more sensitive.
本研究采用基于连接酶检测反应（LDR）与聚合酶链反应（PCR）相结合的新方法，用于检测乙型肝炎病毒（HBV）中低丰度 YIDD 突变体，该方法能够从质粒中 10^8 个野生型拷贝检测到 10^2 个突变体，从临床血清样本中 10^7 个检测到 10^3 个突变体。直接测序和微阵列杂交被选为与该检测方法进行敏感性比较的方法。通过直接测序，在 50 份感染 HBV 的临床标本中发现有 6 份为 YIDD，其余为 YMDD，而通过微阵列杂交，发现 28 份标本同时含有 YMDD 和 YIDD，其余标本只含有 YMDD，通过该检测方法，发现 31 份标本同时含有 YMDD 和 YIDD，其余标本只含有 YMDD，这表明该检测方法相对更敏感。
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