Influence of labeled primer and labeled dUTP assays on the signal intensity of the chip for the detection of HBV gene polymorphism.
- Author:
Da MA
1
;
Huimin WANG
;
Jianlong ZHAO
;
Wanxiang WANG
;
Naizhou GUO
;
Ling JIANG
;
Donglei ZHANG
;
Yue SUN
Author Information
- Publication Type:Journal Article
- MeSH: DNA, Viral; isolation & purification; Fluorescent Dyes; Genome, Viral; Hepatitis B; virology; Hepatitis B virus; genetics; Humans; Polymerase Chain Reaction; methods; Polymorphism, Genetic
- From: Chinese Journal of Experimental and Clinical Virology 2002;16(2):166-167
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUNDTo evaluate the influence of assays with primer labeled with fluorochrome (Cy5) and dUTP labeled with Cy5 on the signal intensity of the chip for detection of hepatitis B virus (HBV) gene polymorphism.
METHODSThe P-region and pre-C/C-region of HBV gene were amplified by polymerase chain reaction (PCR) with Cy5 labeled primer or Cy5 labeled dUTP. The amplicons of the two assays were hybridized with chips, scanned and analyzed by computer software for the detection of HBV gene polymorphism.
RESULTSThe signal intensity of assay with Cy5 labeled dUTP was slightly higher than that of assay with Cy5 labeled primer, but non?specific signal intensity of the assay with Cy5 labeled dUTP was higher. The result of 42 samples showed that there was no significant difference between the two assays, and that both had a good repeatability and CV value (15%-20%).
CONCLUSIONSThe assay with Cy5 labeled primer may replace the assay with Cy5 labeled dUTP as a routine method to detect HBV gene polymorphism, and it is simpler and cheaper.
