The AHV-1 FQ-PCR assay was highly reproducible and linear over a range of eight orders of magnitude from 10 2 to 10 9 copies, allowing a precise calculation of viral DNA load in samples containing a wide range of viral DNA amounts, eliminating the need for sample dilution and minimizing sample handling. The results for intra-and inter-assay precision indicate that both intra-assay and inter-assay CVs were satisfactorily low and the assay is reproducible, even between different batches of reagents used. Probability rather than sample quality variation is the predominant cause of variability at low copy numbers [27].
Conclusion
In conclusion, the FQ-PCR developed in this study is highly specific and sensitive with better parameters than conventional PCR method and is a valuable method for the detection of AHV-1. The method described in this study is especially helpful for high throughput analysis such as evaluating the efficacy of antiviral drugs and experimental vaccines for AHV-1. The research group of authors is currently using this technique to study the AHV-1 distribution characteristics in vaccinated birds and in artificially infected birds. We believe that this method could expedite related AHV-1 research in the AHV-1 viral molecular biology.
Cell, virus and PCR template DNA preparation
Duck embryo fibroblast (DEF) monolayer was incubated at 37°C with 5% CO 2 in tissue culture flasks with Minimal Essential Medium (MEM) that contained 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μg/mL streptomycin.

### Language ###
English

### Assessment ###
Finding out the temperature condition for cell incubation in the laboratory protocol

### Query ###
Any specific temperature needed for duck embryo cell growth?
Any specific temperature needed for duck embryo cell growth?