A novel method based on ligase detection reaction for low abundant YIDD mutants detection in hepatitis B virus.
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.
Xiao
Zhenxian
Z
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.
Xiao
Junxua
J
Jiang
Yuexing
Y
Zhang
Shiyang
S
Yu
Minghui
M
Zhao
Jianlong
J
Wei
Dongzhi
D
Cao
Huimin
H