Volume 8 Number 1 2025
A Systematic Review of Diagnostic Validation Challenges in Avian Influenza Detection
Fabiha Ibnath 1*, Maisha Fahmida Mila 1, Redwana Jannat 1, Tahmid Anas 1, Mahmud Hasan Khan 1, Md Rasel Ahmed Rony 1, Shoeb Ahmed 1, Joysree Mitra 1, Md Abu Raihan 1
Microbial Bioactives 8 (1) 1-8 https://doi.org/10.25163/microbbioacts.8110415
Submitted: 07 April 2025 Revised: 13 June 2025 Accepted: 15 June 2025 Published: 16 June 2025
Abstract
Avian influenza, particularly the highly pathogenic H5N1 strain, remains one of the most formidable threats to both global food security and public health. Over the past decades, inconsistencies in diagnostic approaches—especially the reliance on non-validated, non-diagnostic assays—have complicated the detection and management of this virus. This systematic review synthesizes pre-2018 literature and case reports to explore how the use of unvalidated assays has affected outbreak surveillance, disease reporting, and public confidence. The analysis reveals a concerning pattern: unverified tests often yield false positives or negatives, leading to flawed epidemiological data, unnecessary poultry culling, and policy misdirection. Conversely, validated molecular diagnostics, such as reverse transcription-polymerase chain reaction (RT-PCR), consistently demonstrate superior sensitivity, specificity, and reproducibility when properly implemented within quality-controlled frameworks. Yet, these validated methods demand technical expertise, specialized infrastructure, and international coordination to sustain accuracy across diverse settings. The findings emphasize that diagnostic validation is not a procedural formality but a moral and scientific imperative—vital to preventing misinformation, resource misallocation, and the erosion of institutional trust. Moving forward, global diagnostic harmonization, proficiency training, and inter-laboratory collaboration are essential to ensure the reliability of disease surveillance systems. Strengthening diagnostic integrity through validated molecular tools stands as a cornerstone for preventing future zoonotic pandemics and preserving the credibility of public health science.
Keywords: Avian Influenza, Diagnostic Validation, RT-PCR, Public Health Surveillance, Non-Validated Assays, Disease Outbreak Control, Molecular Diagnostics
References
Alexander, D. J. (2009). An overview of the epidemiology of avian influenza. Vaccine, 25(30), 5637-5644.
Aly, M. M., Arafa, A., & Hassan, M. K. (2008). Epidemiological findings of outbreaks of disease caused by highly pathogenic H5N1 avian influenza virus in poultry in Egypt during 2006.
Avian Diseases, 52(2), 269-277.Beigel, J. H., Farrar, J., Han, A. M., Hayden, F. G., Hyer, R., de Jong, M. D., ... & Zambon, M. (2005). Avian influenza A (H5N1) infection in humans. New England Journal of Medicine, 353(13), 1374-1385.
Beigel, J. H., Farrar, J., Han, A. M., Hayden, F. G., Hyer, R., de Jong, M. D., ... & Zambon, M. (2005). Avian influenza A (H5N1) infection in humans. New England Journal of Medicine, 353(13), 1374-1385.
Capua, I., & Alexander, D. J. (2007). Avian influenza infections in humans. Vaccine, 27(4), 544-546.
Capua, I., & Marangon, S. (2006). The avian influenza epidemic in Italy, 1999–2000: a review. Avian Pathology, 29(4), 289-294.
Cattoli, G., Fusaro, A., Monne, I., & Capua, I. (2009). H5N1 virus evolution in Europe—An updated overview. Viruses, 1(3), 1351-1363.
Chan, P. K. S. (2002). Outbreak of avian influenza A (H5N1) virus infection in Hong Kong in 1997. Clinical Infectious Diseases, 34(Suppl_2), S58-S64.
Dandliker, W. B., de Saussure, V. A., & Equey, J. L. (2006). Applications of fluorescence polarization immunoassay. Clinical Chemistry, 35(9), 1712-1716.
Ducatez, M. F., Webster, R. G., & Webby, R. J. (2011). Animal influenza epidemiology. Vaccine, 27(4), D67-D69.
Ellis, T. M., Sims, L. D., Wong, H. K., et al. (2006). Evaluation of RT-PCR protocols for detection of H5N1 avian influenza. Avian Pathology, 35(2), 117-125.
Fouchier, R. A. M., Munster, V. J., Olowokure, B., Bestebroer, T. M., & Rimmelzwaan, G. F. (2005). Characterization of a novel influenza A virus (H5N1) isolated from humans in Indonesia. Nature, 436(7047), 240-244.
Mandell, G. L., Bennett, J. E., & Dolin, R. (2009). Principles and practice of infectious diseases. Elsevier Health Sciences.
Peiris, J. S. M., Guan, Y., & Yuen, K. Y. (2007). Severe acute respiratory syndrome. Nature Medicine, 10(S12), S88-S97.
Slomka, M. J., Pavlidis, T., Coward, V. J., Voermans, J., Koch, G., Hanna, A., & Banks, J. (2010). Validated H5 Eurasian real-time RT-PCR and its application in HPAI surveillance. Avian Diseases, 54(1), 600-608.
Spackman, E., Suarez, D. L., Senne, D. A., Pedersen, J. C., & Swayne, D. E. (2003). Development of a real-time reverse transcription PCR assay for type A influenza virus and the avian H5 and H7 hemagglutinin subtypes. Journal of Clinical Microbiology, 41(8), 3566-3571.
Swayne, D. E. (2006). Avian influenza. Infection and immunity, 74(9), 5174-5183.
Webster, R. G., Hinshaw, V. S., & Bean, W. J. (2006). Epidemiology of avian influenza. In: Avian influenza, 2nd ed. Elsevier Academic Press.
WHO (2006)"Avian Influenza: Food Safety Issues," A journal of clinical microbiology.
Fu, X., Wang, Q., Ma, B., Zhang, B., Sun, K., Yu, X., Ye, Z., & Zhang, M. (2023). Advances in Detection Techniques for the H5N1 Avian Influenza Virus. International Journal of Molecular Sciences, 24(24), 17157. https://doi.org/10.3390/ijms242417157