[Establishment and evaluation of the method for detecting HBV DNA in serum using HBV DNA probe labeled directly by alkaline phosphatase].
OBJECTIVE
To establish a sensitive and specific technique for detecting HBV DNA in serum using HBV DNA probe labeled directly by alkaline phosphatase (AlkPhos Direc probe).


METHODS
The probe that purified HBV DNA sequence was labeled directly by alkaline phosphatase and chemiluminescent substrate CDP-star for AP was used in the hybridization assay. HBV DNA was detected by autoradiography on the film. The test compared the chemiluminescen dot blot hybridization assay for 80 samples with digoxigenin-labeled HBV DNA probe detective method. The correlation of 70 samples test results between fluorescent quantitative HBV DNA PCR method and dot blot hybridization assay by AlkPhos Direc probe was analysed.


RESULTS
The sensitivity of the probe labeled directly by alkaline phosphatase was 10pg at least. The coincidence was 100% compared with digoxigenin-labeled HBV DNA probe detection. A correlation coefficient of HBV DNA quantitative results between fluorescent quantitative HBV DNA PCR (QPCR) method and dot blot hybridization assay by AlkPhos Direc probe was 0.98 (P<0.01).


CONCLUSIONS
The method detecting HBV DNA in serum by HBV DNA AlkPhos Direc probe is sensitive and specific. The results between two methods with AlkPhos Direc and digoxigenin-labeled HBV DNA probe are coincident completely. The correlation of HBV DNA quantitative results between fluorescent QPCR method and dot blot hybridization assay by AlkPhos Direc probe is satisfactory.
Chen
Yaxi
Y
Institute for Viral Hepatitis, Chongqing University of Medical Sciences, Chongqing 400010, China.
Huang
Ailong
A
Qi
Zhenyuan
Z
Shan
Youlan
Y
Sun
Hang
H
D000469:Alkaline Phosphatase / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D015342:DNA Probes / Q000737:chemistry / Q000235:genetics; D004279:DNA, Viral / Q000097:blood* / Q000235:genetics; D006509:Hepatitis B / Q000097:blood* / Q000175:diagnosis / Q000821:virology; D006515:Hepatitis B virus / Q000235:genetics*; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods* / Q000592:standards; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity
D015342:DNA Probes; D004279:DNA, Viral; D000469:Alkaline Phosphatase