[Preparation and application of Epstein-Barr virus genome-wide genechip].
OBJECTIVE
To prepare an Epstein-Barr virus (EBV) microarray using known and predicted EBV-coded genes as the cDNA probes to detect the EBV gene expression in nasopharyngeal carcinoma (NPC) tissues.


METHODS
The EBV gene probes were amplified by PCR using a pair of primers designed in both sides of the multiple clone site (MCS) of the T/A vector. After purification of the PCR products, 85 EBV genes and 8000 human genes were printed onto the same slide as the detection chip consisting of both EBV and human genes. This genechip was used to detect the differential gene expression in NPC and non-cancerous nasopharynx (NP) tissues.


RESULTS
Detection of the human gene expression profile using the prepared genechip resulted in the identification of numerous human genes in the tissue specimens. Some EBV genes were also detected in the tissues using the genechip, but the signals of the genes appeared rather weak without distinctly visible fluorescence, and were not comparable to the strong signal intensities of the human genes.


CONCLUSIONS
The EBV microarray, though constructed successfully, can not meet the needs for clinical application due to the limited detection sensitivity and the relative small quantity of EBV gene expression in NPC samples. Further improvements of the research methods are warranted.
Cancer Research Institute, Key Lab for Transcriptomics and Proteomics of Human Fatal Diseases supported by Ministry of Education and Guangdong Province, Southern Medical University, Guangzhou, China. fangweiyi1975@yahoo.com.cn
Wei-yi
WY
Fang
Wen-ling
WL
Zheng
Wen-li
WL
Ma
Zhen
Z
Liu
Xin
X
Li
Kai-tai
KT
Yao
D002294:Carcinoma, Squamous Cell / Q000821:virology*; D020869:Gene Expression Profiling; D016679:Genome, Viral / Q000235:genetics; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D009303:Nasopharyngeal Neoplasms / Q000821:virology*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*