DIGOXYGENIN-LABELED DNA-PROBE - A RAPID NONRADIOACTIVE METHOD FOR HEPATITIS-B VIRUS-DNA DETECTION IN SERUM

被引:0
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作者
BUTI, M
JARDI, R
RODRIGUEZFRIAS, F
ARRANZ, JA
CASACUBERTA, JM
SANSEGUNDO, B
PUIGDOMENECH, P
ESTEBAN, R
GUARDIA, J
机构
[1] UNIV AUTONOMA BARCELONA, HOSP VALLE HEBRON DEPT INTERNAL MED, LIVER UNIT, E-08035 BARCELONA, SPAIN
[2] UNIV AUTONOMA BARCELONA, HOSP VALLE HEBRON, DEPT BIOCHEM, LIVER UNIT, E-08035 BARCELONA, SPAIN
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中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The sensitivity and specificity of two non-radioactive spot hybridization assays for hepatitis B virus DNA (HBV-DNA) using biotin and digoxygenin-labelled DNA probes were investigated in parallel in 122 serum samples from patients with chronic hepatitis B and 50 controls. The results were compared with an isotopic technique using a P-32-labelled probe. HBV-DNA was detected in 56 (80%) out of 70 hepatitis B "e" antigen (HBeAg)-positive cases and in 4 (8%) out of 52 antibody to hepatitis B "e" antigen (anti-HBe)-positive cases using the digoxygenin or P-32-labelled probes. No false positives were found with either method. Using the biotin-labelled probe, 16% of sera gave discordant results, which were considered to be false positive. The time required for detection of serum HBV-DNA was 2 hours for the non-radioactive probes and 16 hours for the isotopic probes. This study suggests that the digoxygenin-labelled probe for detection of HBV-DNA is the most rapid and sensitive method for routine diagnosis of viral replication in clinical laboratories.
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页码:731 / 735
页数:5
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