Abstract：
:To develop a quantitative assay for universal detection of duck hepatitis B virus (DHBV) DNA, a Taqman real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) assay was developed using primers and probes based on genomic sequences located at nucleotide 241-414 of the DHBV Core region which possesses the highest homology among the 44 DHBV genomes available in Genbank. The DHBV Core gene cloned in pGEM-T was used to generate DHBV DNA standard. The assay had a lowest detection limit of 10(3) copies/ml and a good linear standard curve (Y=-3.989X+49.086, r(2)=0.9993) over a wide range of input DHBV DNA (10(3) to 10(10) copies/ml). The standard deviation of intra- and inter-assay was 0.01-0.06 and 0.05-0.16, respectively, and the coefficient of variation was 1.3-1.8%. The specificity of the assay was validated using duck hepatitis virus type 1, hepatitis B virus, and E. coli DNA. Comparison of ABI 7300 and Bio-Rad iQ5 PCR instruments yielded highly consistent results. The assay showed a positive rate of 63.8% (51/80) DHBV DNA in peripheral blood and liver tissue from ducks from Xi'an, China. The FQ-PCR developed is highly sensitive, specific, reproducible and versatile, and may be used to universally detect DHBV DNA of different DHBV strains.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Wang Y,Li Y,Yang C,Hui L,Han Q,Ma L,Wang Q,Yang G,Liu Zdoi
10.1016/j.jviromet.2013.03.025subject
Has Abstractpub_date
2013-07-01 00:00:00pages
41-7issue
1eissn
0166-0934issn
1879-0984pii
S0166-0934(13)00101-8journal_volume
191pub_type
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