previous page   Pak Vet J, xxxx, xx(x): xxx   next page
Development of Taqman Real-time Fluorescent Quantitative PCR for Rapid Detection and differentiation between DHAV-1 and DHAV-3 in Duck Farming
Ahmed Abd Elhalem Mohamed1, Wesam H. Mady1, Sabry E. Omar2, Lamya A.F. Atteya2, Mariam Abdulaziz Alkhateeb3*, Amin A. Al-Doaiss4, Ohud Saleh5, Nada Alhazmi6,7, Ashwaq M. Al-Nazawi8, Dalia Said1, and Nahed Yehia1

1Reference Laboratory for Veterinary Quality Control on Poultry Production (RLQP), Animal Health Research Institute, Agriculture Research Center (ARC), Giza 12618, Egypt; 2Animal Health Research Institute, Banha Branch, Agricultural Research Center, Giza, Egypt; 3Department of Biology College of Science Princess Nourah bint Abdulrahman University, P.O.BOX 84428, Riyadh 11671, Saudi Arabia; 4Biology Department, College of Science, King Khalid University, P.O. Box 9004, Abha 61413, Saudi Arabia; 5Department of Biochemistry, College of Science, University of Jeddah, Jeddah 21959, Saudi Arabia; 6Department of Basic Sciences, College of Science and Health Professions, King Saud bin Abdulaziz University for Health Sciences, Jeddah 11481, Saudi Arabia; 7King Abdullah International Medical Research Center, Jeddah, Saudi Arabia; 8Department of Epidemiology, Faculty of Public Health and Tropical Medicine, Jazan University, Jazan 82726, Saudi Arabia
*Corresponding author:


The duck industry is at high risk from duck viral hepatitis (DVH) and has the potential to cause substantial financial losses because of the high mortality rates observed in duck farming, even with continuous breeder duck flock vaccination. Among the different etiological agents of DVH, duck hepatitis A virus type-1 (DHAV-1) is the most common followed by DHAV-2 and DHAV-3. Although DHAV-I is more common and pathogenic, DHAV-3 has just emerged from duck farms in North Egypt, thus there's a pressing need to find a way to detect both DHAV-1 and DHAV-3 rapidly and simultaneously using real-time qPCR. To assess and compare the sensitivity of the real time reverse transcriptase PCR (rRT-PCR) technique for the detection of DHAV-3 and DHAV-1, dilution range of titrated DHAV-1 and DHAV-3 reference strains from 107.2 and 106 EID50/ml to 1EID50, was implemented, respectively. The results of the current study confirmed that the rRT-PCR assay's had lowest detection limit for DHAV-1 and DHAV-3 was 102.2 and 102 EID50/ml, respectively, and it is ten-fold higher than RT-PCR. The rRT-PCR was highly specific to DHAV-1 and DHAV-3, as other avian diseases and nucleic acid isolated from samples that tested negative for DHAV. When examining clinical samples for rRT-PCR, the diagnostic sensitivity was better than the RT-PCR. It detected 25 out of 40 clinical suspected samples but the RT-PCR detected only 15 out of 40 clinical suspected samples. In conclusion, the assay may be used as an efficient, rapid, sensitive, specific, and focused molecular diagnostic technique for detection and epidemiological investigations of DVH caused by both DHAV-1 and DHAV-3.

To Cite This Article: Abdelhalim AM, Mady WH, Omar SE, Atteya LAF, Alkhateeb MA, Al-Doaiss AA, Saleh O, Alhazmi N, Al-Nazawi AM., Said D and Yehia N, 2024. Development of Taqman real-time fluorescent quantitative PCR for rapid detection and differentiation between DHAV-1 and DHAV-3 in duck farming. Pak Vet J.


ISSN 0253-8318 (Print)
ISSN 2074-7764 (Online)

SCImago Journal & Country Rank