Development of TaqMan MGB fluorescent real-time PCR assay for the detection of anatid herpesvirus 1.
BACKGROUND
Anatid herpesvirus 1 (AHV-1) is an alphaherpesvirus associated with latent infection and mortality in ducks and geese and is currently affecting the world-wide waterfowl production severely. Here we describe a fluorescent quantitative real-time PCR (FQ-PCR) method developed for fast measurement of AHV-1 DNA based on TaqMan MGB technology.


RESULTS
The detection limit of the assay was 1 x 101 standard DNA copies, with a sensitivity of 2 logs higher than that of the conventional gel-based PCR assay targeting the same gene. The real-time PCR was reproducible, as shown by satisfactory low intra-assay and inter-assay coefficients of variation.


CONCLUSIONS
The high sensitivity, specificity, simplicity and reproducibility of the AHV-1 fluorogenic PCR assay, combined with its wide dynamic range and high throughput, make this method suitable for a broad spectrum of AHV-1 etiologically related application.
Guo
Yufei
Y
Avian Disease Research Center, Sichuan Agricultural University, Yaan, PR China. gyf02@163.com
Cheng
Anchun
A
Wang
Mingshu
M
Shen
Chanjuan
C
Jia
Renyong
R
Chen
Shun
S
Zhang
Na
N
D018138:Alphaherpesvirinae / Q000235:genetics* / Q000302:isolation & purification*; D000818:Animals; D001715:Bird Diseases / Q000175:diagnosis* / Q000821:virology; D004279:DNA, Viral / Q000235:genetics; D004372:Ducks / Q000821:virology; D005777:Geese / Q000821:virology; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D011201:Poultry Diseases / Q000175:diagnosis* / Q000821:virology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D004279:DNA, Viral