Microarrays for the detection of HBV and HDV.
The increasing pace of development in molecular biology during the last decade has had a direct effect on mass testing and diagnostic applications, including blood screening. We report the model Microarray that has been developed for Hepatitis B virus (HBV) and Hepatitis D virus (HDV) detection. The specific primer pairs of PCR were designed using the Primer Premier 5.00 program according to the conserved regions of HBV and HDV. PCR fragments were purified and cloned into pMD18-T vectors. The recombinant plasmids were extracted from positive clones and the target gene fragments were sequenced. The DNA microarray was prepared by robotically spotting PCR products onto the surface of glass slides. Sequences were aligned, and the results obtained showed that the products of PCR amplification were the required specific gene fragments of HBV, and HDV. Samples were labeled by Restriction Display PCR (RD-PCR). Gene chip hybridizing signals showed that the specificity and sensitivity required for HBV and HDV detection were satisfied. Using PCR amplified products to construct gene chips for the simultaneous clinical diagnosis of HBV and HDV resulted in a quick, simple, and effective method. We conclude that the DNA microarray assay system might be useful as a diagnostic technique in the clinical laboratory. Further applications of RD-PCR for the sample labeling could speed up microarray multi-virus detection.
Institute of Molecular Oncology, General Hospital of Guangzhou Command, Guangzhou 510010, P. R. China.
Sun
S
Zhaohui
Zheng
Z
Wenling
Zhang
Z
Bao
Shi
S
Rong
Ma
M
Wenli
D001483:Base Sequence; D015342:DNA Probes / Q000235:genetics; D005456:Fluorescent Dyes / Q000737:chemistry; D006509:Hepatitis B / Q000175:diagnosis*; D006515:Hepatitis B virus / Q000235:genetics / Q000302:isolation & purification*; D003699:Hepatitis D / Q000175:diagnosis*; D003698:Hepatitis Delta Virus / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012371:Robotics
D015342:DNA Probes; D005456:Fluorescent Dyes