Related Experiment Video
Updated: Jul 19, 2026
Highly Sensitive Assay for Measurement of Arenavirus-cell Attachment
Published on: March 2, 2016
[Establishing a real-time PCR assay for anatid herpesvirus 1]
Fa-long Yang1, Wen-xiang Jia, Yi Xie
1Department of Microbiology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Objectives:
To develop a real-time quantitative PCR assay to detect duck plague virus (DPV) for the rapid diagnosis of DPV infection, the investigation of its nosogenesis and the screening of effective antiviral drugs.
Methods:
The primer and probe were designed according to the gene sequence of DPV DNA polymerase gene. To establish a standard curve, a plasmid containing 125 bp PCR product was constructed and severed as a positive control. The TaqMan based real-time PCR method was adopted and compared with the traditional PCR approach in sensitivity, reliability and specificity.
Results:
A good linear correlation was demonstrated in the standard curve for the real-time PCR assay within the range from 2.3 x 10 to 2.3 x 10(5) copies. A minimum of 23 positive plasmids could be detected, indicating a good sensitivity of the assay. The coefficients of variance (CVs) were 1.22-6.69 and 2.09-8.84 for the intra-assay and inter-assay tests respectively, indicating a good reliability. No amplification products were found for DNA from other pathogens, indicating a good specificity. The comparative study proved that the TaqMan technology was much more sensitive than traditional PCR assay.
Conclusion:
The real-time quantitative PCR assay for DPV DNA has good sensitivity, specificity and reliability.
