Detection of hepatitis B virus DNA by real-time PCR using TaqMan-MGB probe technology.
World J Gastroenterol
; 11(4): 508-10, 2005 Jan 28.
Article
de En
| MEDLINE
| ID: mdl-15641135
ABSTRACT
AIM:
To develop a real-time PCR for detecting hepatitis B virus (HBV) DNA based on TaqMan technology using a new MGB probe.METHODS:
Plasmid containing the sequence of X gene (1414-1744 nt) was constructed as HBV-DNA standard for quantitative analysis. A TaqMan-MGB probe between primers for amplification was designed to detect PCR products. The interested sequence contained in the plasmid and in clinical specimens was quantitatively measured.RESULTS:
The detection limit of the assay for HBV DNA was 1 genome equivalent per reaction. A linear standard curve was obtained between 10(0) and 10(9) DNA copies/reaction (r>0.990). None of the negative control samples showed false-positive reactions in duplicate. HBV DNA was detected in 100% (50/50) of HBV patients with HbeAg, and in 72.0% (36/50) with HBsAg, HBeAb and HBcAb. The coefficient of variation for both intra- and inter-experimental variability demonstrated high reproducibility and accuracy.CONCLUSION:
Real-time PCR based on TaqMan-MGB probe technology is an excellent method for detection of HBV DNA.
Texte intégral:
1
Collection:
01-internacional
Base de données:
MEDLINE
Sujet principal:
Virus de l'hépatite B
/
Réaction de polymérisation en chaîne
/
Hépatite B chronique
/
TAQ polymerase
Type d'étude:
Diagnostic_studies
Limites:
Humans
Langue:
En
Journal:
World J Gastroenterol
Sujet du journal:
GASTROENTEROLOGIA
Année:
2005
Type de document:
Article
Pays d'affiliation:
Chine