Diagnostic accuracy of loop-mediated isothermal amplification (LAMP) assays for dengue virus detection in serum: A systematic review and meta-analysis
DOI:
https://doi.org/10.61511/phraj.v3i2.2026.2247Keywords:
LAMP, dengeu virus, diagnostic accuracyAbstract
Background: Rapid and accurate diagnosis of dengue virus (DENV) infection remains a challenge in endemic areas. Current gold standard methods have several limitations and are often unsuitable for resource-limited settings. Loop-mediated isothermal amplification (LAMP) offers a rapid, cost-effective, and field-adaptable alternative. This meta-analysis aimed to evaluate the diagnostic accuracy of LAMP for detecting DENV infection in human serum samples. Methods: A comprehensive literature search was conducted in PubMed, Scopus, Taylor & Francis, and Wiley databases up to July 2025. Pooled sensitivity, specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR), diagnostic odds ratio (DOR), and area under the curve (AUC) were calculated to assess the diagnostic performance of LAMP. Meta-DiSC 1.4 was used for analysis, and methodological quality was evaluated using the QUADAS-2 tool. Meta-regression was performed to explore potential sources of heterogeneity. Findings: Five studies involving 807 samples were included in this meta-analysis. The pooled meta-analysis results were as follows: sensitivity 0.83 (95% CI: 0.80–0.85), specificity 0.95 (95% CI: 0.91–0.98), PLR 14.31 (95% CI: 7.82–26.20), NLR 0.15 (95% CI: 0.07–0.31), and DOR 103.30 (95% CI: 23.13–461.42). The summary of AUC was 0.9633, indicating good diagnostic accuracy. Meta-regression showed no significant effect of study design, sample size, geographic region, cross-reactivity testing, or reference standard on diagnostic accuracy. Conclusion: LAMP provides a highly accurate and reliable method for DENV detection in human serum, suitable for both clinical and field use. Its routine implementation may improve dengue outbreak management and surveillance in endemic areas. Novelty/Originality of this article: This is the first meta-analysis to comprehensively evaluate the diagnostic accuracy of LAMP for DENV detection in human serum. By synthesizing evidence from multiple studies, it provides stronger statistical power than individual reports and highlights the robustness of LAMP across diverse settings.
References
Aborode, A. T., Adesola, R. O., Scott, G. Y., Arthur-Hayford, E., Otorkpa, O. J., Kwaku, S. D., Elebesunu, E. E., Nibokun, E. O., Aruorivwooghene, I. J., Bakre, A. A., Ogundijo, O. A., Banwo, O. G., Ige, O., Adelakun, I. O., Onifade, I. A., Ogungbemi, S. E., Dosunmu, B. T., Ayando, O. D., Idowu, N., … Jimoh, O. O. (2025). Bringing lab to the field: Exploring innovations in point-of-care diagnostics for the rapid detection and management of tropical diseases in resource-limited settings. Advances in Biomarker Sciences and Technology, 7, 28–43. https://doi.org/10.1016/j.abst.2025.01.001
Alhamid, G., Tombuloglu, H., & Al-Suhaimi, E. (2023). Development of loop-mediated isothermal amplification (LAMP) assays using five primers reduces the false-positive rate in COVID-19 diagnosis. Scientific Reports, 13. https://doi.org/10.1038/s41598-023-31760-z
Arruda, V. de O., Filho, L. R. G., & Neves, A. F. (2024). Aptamer-associated colorimetric reverse transcription loop-mediated isothermal amplification assay for detection of dengue virus. Microbiology Spectrum, 12(9). https://doi.org/10.1128/spectrum.03583-23
Artika, I. M., Dewi, Y. P., Nainggolan, I. M., Siregar, J. E., & Antonjaya, U. (2022). Real-Time Polymerase Chain Reaction: Current Techniques, Applications, and Role in COVID-19 Diagnosis. Genes, 13(12). https://doi.org/10.3390/genes13122387
Berba, R. P., Santos, J. A. P., Flores, K. M. G., Tonga, A. dela, Galarion, M. J., Carascal, M. B., Camacho, A., Aguirre, C. A., Lecciones, J. A., & Gonzales, M. L. M. (2021). Clinical Validation Of Biotek-Mtm Dengue Aqua Kit In The Diagnosis Of Dengue Infections In The Philippines. The Southeast Asian Journal of Tropical Medicine and Public Health, 52(1). https://journal.seameotropmednetwork.org/index.php/ jtropmed/article/view/385
Bonney, J. H. K., Pratt, D., Ofori, M., Hayashi, T., Abankwa, A., Awuku-Larbi, Y., Kumordjie, S., Agbodzi, B., Salisu, M., Mante, A. A. O., Bour, S., Eshun, M., Amaning, J. N. D. A., Ketorwoley, P., Enimil, N., Koomson, J., Stephens, G., Asiedu-Bekoe, F., Laryea, D., … Suzuki, T. (2024). Molecular detection of dengue virus from febrile patients in Ghana. BMC Infectious Diseases, 24(1). https://doi.org/10.1186/s12879-024-10289-0
Caraballo, G. I., Rosales, R., Viettri, M., Castillo, J. M., Cruz, R., Ding, S., Greenberg, H. B., & Ludert, J. E. (2022). The Dengue Virus Nonstructural Protein 1 (NS1) Interacts with the Putative Epigenetic Regulator DIDO1 to Promote Flavivirus Replication in Mosquito Cells. Journal of Virology, 96(12). https://doi.org/10.1128/jvi.00704-22
Cassedy, A., Parle-McDermott, A., & O’Kennedy, R. (2021). Virus Detection: A Review of the Current and Emerging Molecular and Immunological Methods. Frontiers in Molecular Biosciences, 8, 637559. https://doi.org/10.3389/fmolb.2021.637559
CDC. (2025, June 20). Clinical Testing Guidance for Dengue. Dengue. https://www.cdc.gov/dengue/hcp/diagnosis-testing/index.html
Chan, K. R., Ismail, A. A., Thergarajan, G., Raju, C. S., Yam, H. C., Rishya, M., & Sekaran, S. D. (2022). Serological cross-reactivity among common flaviviruses. Frontiers in Cellular and Infection Microbiology, 12. https://doi.org/10.3389/fcimb.2022.975398
Chen, P.-K., Chang, J.-H., Ke, L.-Y., Kao, J.-K., Chen, C.-H., Yang, R.-C., Yoshimura, T., Ito, E., & Tsai, J.-J. (2023). Advanced Detection Method for Dengue NS1 Protein Using Ultrasensitive ELISA with Thio-NAD Cycling. Viruses, 15(9), 1894. https://doi.org/10.3390/v15091894
Ciotti, M., Nicolai, E., & Pieri, M. (2024). Development and optimization of diagnostic assays for infectious diseases. LabMed Discovery, 1(2), 100032. https://doi.org/10.1016/j.lmd.2024.100032
Dauner, A. L., Mitra, I., Gilliland, T., Seales, S., Pal, S., Yang, S.-C., Guevara, C., Chen, J.-H., Liu, Y.-C., Kochel, T. J., & Wu, S.-J. L. (2015). Development of a pan-serotype reverse transcription loop-mediated isothermal amplification assay for the detection of dengue virus. Diagnostic Microbiology and Infectious Disease, 83(1), 30–36. https://doi.org/10.1016/j.diagmicrobio.2015.05.004
Dietrich, V. C., Costa, J. M. C., Oliveira, M. M. G. L., Aguiar, C. E. O., Silva, L. G. de O., Luz, M. S., Lemos, F. F. B., & Melo, F. F. de. (2025). Pathogenesis and clinical management of arboviral diseases. World Journal of Virology, 14(1). https://doi.org/10.5501/wjv.v14.i1.100489
Dorta-Gorrín, A., Navas-Méndez, J., Gozalo-Margüello, M., Miralles, L., & García-Hevia, L. (2023). Detection of SARS-CoV-2 Based on Nucleic Acid Amplification Tests (NAATs) and Its Integration into Nanomedicine and Microfluidic Devices as Point-of-Care Testing (POCT). International Journal of Molecular Sciences, 24(12), 10233. https://doi.org/10.3390/ijms241210233
Garg, N., Ahmad, F. J., & Kar, S. (2022). Recent advances in loop-mediated isothermal amplification (LAMP) for rapid and efficient detection of pathogens. Current Research in Microbial Sciences, 3. https://doi.org/10.1016/j.crmicr.2022.100120
Glökler, J., Lim, T. S., Ida, J., & Frohme, M. (2021). Isothermal amplifications – a comprehensive review on current methods. Critical Reviews in Biochemistry and Molecular Biology, 56(6), 543–586. https://doi.org/10.1080/10409238.2021.1937927
Gomes Torres, A. C. M. B., Mathias, C., Baal, S. C. S., Kohler, A. F., Cunha, M. L., & Blanes, L. (2024). Advancements in LAMP-Based Diagnostics: Emerging Techniques and Applications in Viral Detection with a Focus on Herpesviruses in Transplant Patient Management. International Journal of Molecular Sciences, 25(21). https://doi.org/10.3390/ijms252111506
Gunasegar, S., & Neela, V. K. (2021). Evaluation of diagnostic accuracy of loop-mediated isothermal amplification method (LAMP) compared with polymerase chain reaction (PCR) for Leptospira spp. in clinical samples: A systematic review and meta-analysis. Diagnostic Microbiology and Infectious Disease, 100(3). https://doi.org/10.1016/j.diagmicrobio.2021.115369
Gwee, S. X. W., St John, A. L., Gray, G. C., & Pang, J. (2021). Animals as potential reservoirs for dengue transmission: A systematic review. One Health, 12. https://doi.org/10.1016/j.onehlt.2021.100216
Han, Y., Sun, X., & Cai, S. (2025). Advances in Research on Isothermal Signal Amplification Mediated MicroRNA Detection of Clinical Samples: Application to Disease Diagnosis. Biosensors, 15(6), 395. https://doi.org/10.3390/bios15060395
Hanifehpour, H., Ashrafi, F., Siasi, E., & Fallahi, S. (2024). Evaluation and comparison of one-step real-time PCR and one-step RT-LAMP methods for detection of SARS-CoV-2. BMC Infectious Diseases, 24(1), 679. https://doi.org/10.1186/s12879-024-09574-9
Harapan, H., Michie, A., Sasmono, R. T., & Imrie, A. (2020). Dengue: A Minireview. Viruses, 12(8), Article 8. https://doi.org/10.3390/v12080829
Harshitha, R., & Arunraj, D. R. (2021). Real-time quantitative PCR: A tool for absolute and relative quantification. Biochemistry and Molecular Biology Education, 49(5), 800–812. https://doi.org/10.1002/bmb.21552
Hegde, S. S., & Bhat, B. R. (2022). Dengue detection: Advances and challenges in diagnostic technology. Biosensors and Bioelectronics: X, 10. https://doi.org/10.1016/j.biosx.2021.100100
Hodinka, R. L., & Kaiser, L. (2013). Is the Era of Viral Culture Over in the Clinical Microbiology Laboratory? Journal of Clinical Microbiology, 51(1), 2–8. https://doi.org/10.1128/JCM.02593-12
Hurtado-Gómez, L., Escorcia-Lindo, K., Rosero, J. S., Solano Llanos, N., Barrios Sánchez, C., Díaz Pérez, A., Díaz-Olmos, Y., García, J., Bello-Lemus, Y., Pacheco-Londoño, L. C., Acosta Hoyos, A. J., & Pacheco-Lugo, L. A. (2025). Development and Validation of a Combined RT-LAMP Assay for the Rapid and Sensitive Detection of Dengue Virus in Clinical Samples from Colombia. Diagnostics, 15(5), 570. https://doi.org/10.3390/diagnostics15050570
Kabir, M. A., Zilouchian, H., Younas, M. A., & Asghar, W. (2021). Dengue Detection: Advances in Diagnostic Tools from Conventional Technology to Point of Care. Biosensors, 11(7), 206. https://doi.org/10.3390/bios11070206
Kellerhuis, B. E., Jenniskens, K., Schuit, E., Hooft, L., Moons, K. G. M., & Reitsma, J. B. (2025). Drivers of bias in diagnostic test accuracy estimates when using expert panels as a reference standard: A simulation study. BMC Medical Research Methodology, 25, 106. https://doi.org/10.1186/s12874-025-02557-7
Khan, M. B., Yang, Z.-S., Lin, C.-Y., Hsu, M.-C., Urbina, A. N., Assavalapsakul, W., Wang, W.-H., Chen, Y.-H., & Wang, S.-F. (2023). Dengue overview: An updated systemic review. Journal of Infection and Public Health, 16(10), 1625–1642. https://doi.org/10.1016/j.jiph.2023.08.001
Kok, B. H., Lim, H. T., Lim, C. P., Lai, N. S., Leow, C. Y., & Leow, C. H. (2022). Dengue virus infection – a review of pathogenesis, vaccines, diagnosis and therapy. Virus Research, 324, 199018. https://doi.org/10.1016/j.virusres.2022.199018
Kularatne, S. A., & Dalugama, C. (2022). Dengue infection: Global importance, immunopathology and management. Clinical Medicine, 22(1), 9–13. https://doi.org/10.7861/clinmed.2021-0791
Kumar, S., Sharma, S., Bhardwaj, N., Pande, V., Savargaonkar, D., & Anvikar, A. R. (2021). Advanced Lyophilised Loop Mediated Isothermal Amplification (L-LAMP) based point of care technique for the detection of dengue virus. Journal of Virological Methods, 293, 114168. https://doi.org/10.1016/j.jviromet.2021.114168
Kutsuna, S., Saito, S., & Ohmagari, N. (2020). Simultaneous diagnosis of dengue virus, Chikungunya virus, and Zika virus infection using a new point-of-care testing (POCT) system based on the loop-mediated isothermal amplification (LAMP) method. Journal of Infection and Chemotherapy, 26(12), 1249–1253. https://doi.org/10.1016/j.jiac.2020.07.001
Lambrechts, L., Reiner, R. C., Briesemeister, M. V., Barrera, P., Long, K. C., Elson, W. H., Vizcarra, A., Astete, H., Bazan, I., Siles, C., Vilcarromero, S., Leguia, M., Kawiecki, A. B., Perkins, T. A., Lloyd, A. L., Waller, L. A., Kitron, U., Jenkins, S. A., Hontz, R. D., … Morrison, A. C. (2023). Direct mosquito feedings on dengue-2 virus-infected people reveal dynamics of human infectiousness. PLOS Neglected Tropical Diseases, 17(9). https://doi.org/10.1371/journal.pntd.0011593
Mathur, M. B., & VanderWeele, T. J. (2021). Meta-regression methods to characterize evidence strength using meaningful-effect percentages conditional on study characteristics. Research Synthesis Methods, 12(6), 731–749. https://doi.org/10.1002/jrsm.1504
Melo, C. L., Mageste, L. R., Guaraldo, L., Paula, D. P., & Wakimoto, M. D. (2024). Use of Digital Tools in Arbovirus Surveillance: Scoping Review. Journal of Medical Internet Research, 26(1), e57476. https://doi.org/10.2196/57476
Nanaware, N., Banerjee, A., Mullick Bagchi, S., Bagchi, P., & Mukherjee, A. (2021). Dengue Virus Infection: A Tale of Viral Exploitations and Host Responses. Viruses, 13(10), 1967. https://doi.org/10.3390/v13101967
Özay, B., & McCalla, S. E. (2021). A review of reaction enhancement strategies for isothermal nucleic acid amplification reactions. Sensors and Actuators Reports, 3, 100033. https://doi.org/10.1016/j.snr.2021.100033
Page, M. J., McKenzie, J. E., Bossuyt, P. M., Boutron, I., Hoffmann, T. C., Mulrow, C. D., Shamseer, L., Tetzlaff, J. M., Akl, E. A., Brennan, S. E., Chou, R., Glanville, J., Grimshaw, J. M., Hróbjartsson, A., Lalu, M. M., Li, T., Loder, E. W., Mayo-Wilson, E., McDonald, S., … Moher, D. (2021). The PRISMA 2020 statement: An updated guideline for reporting systematic reviews. BMJ, 372. https://doi.org/10.1136/bmj.n71
Panno, S., Matić, S., Tiberini, A., Caruso, A. G., Bella, P., Torta, L., Stassi, R., & Davino, S. (2020). Loop Mediated Isothermal Amplification: Principles and Applications in Plant Virology. Plants, 9(4), 461. https://doi.org/10.3390/plants9040461
Papadakis, G., Pantazis, A. K., Fikas, N., Chatziioannidou, S., Tsiakalou, V., Michaelidou, K., Pogka, V., Megariti, M., Vardaki, M., Giarentis, K., Heaney, J., Nastouli, E., Karamitros, T., Mentis, A., Zafiropoulos, A., Sourvinos, G., Agelaki, S., & Gizeli, E. (2022). Portable real-time colorimetric LAMP-device for rapid quantitative detection of nucleic acids in crude samples. Scientific Reports, 12, 3775. https://doi.org/10.1038/s41598-022-06632-7
Park, J.-W. (2022). Principles and Applications of Loop-Mediated Isothermal Amplification to Point-of-Care Tests. Biosensors, 12(10), 857. https://doi.org/10.3390/bios12100857
Paul, J. K., Azmal, M., Alam, T., Talukder, O. F., & Ghosh, A. (2025). Comprehensive analysis of intervention and control studies for the computational identification of dengue biomarker genes. PLOS Neglected Tropical Diseases, 19(3). https://doi.org/10.1371/journal.pntd.0012914
Picot, S., Cucherat, M., & Bienvenu, A.-L. (2020). Systematic review and meta-analysis of diagnostic accuracy of loop-mediated isothermal amplification (LAMP) methods compared with microscopy, polymerase chain reaction and rapid diagnostic tests for malaria diagnosis. International Journal of Infectious Diseases: IJID: Official Publication of the International Society for Infectious Diseases, 98, 408–419. https://doi.org/10.1016/j.ijid.2020.07.009
Pourzangiabadi, M., Najafi, H., Fallah, A., Goudarzi, A., & Pouladi, I. (2025). Dengue virus: Etiology, epidemiology, pathobiology, and developments in diagnosis and control – A comprehensive review. Infection, Genetics and Evolution, 127. https://doi.org/10.1016/j.meegid.2024.105710
Robert, S., Diagbouga, P. S., Djibougou, A. D., Guy, D., Bagnall, R., & Ravel, F. (2025). Dengue diagnosis and impact on clinical management: A literature review. PLOS Neglected Tropical Diseases, 19(6). https://doi.org/10.1371/journal.pntd.0013196
Santoso, M. S., Nara, M. B. R., Nugroho, D. K., Yohan, B., Purnama, A., Boro, A. M. B., Hayati, R. F., Gae, E. P., Denis, D., Rana, B., Hibberd, M. L., & Sasmono, R. T. (2025). Investigation of severe dengue outbreak in Maumere, East Nusa Tenggara, Indonesia: Clinical, serological, and virological features. PLOS ONE, 20(2). https://doi.org/10.1371/journal.pone.0317854
Santoso, M. S., Yohan, B., Denis, D., Hayati, R. F., Haryanto, S., Trianty, L., Noviyanti, R., Hibberd, M. L., & Sasmono, R. T. (2020). Diagnostic accuracy of 5 different brands of dengue virus non-structural protein 1 (NS1) antigen rapid diagnostic tests (RDT) in Indonesia. Diagnostic Microbiology and Infectious Disease, 98(2). https://doi.org/10.1016/j.diagmicrobio.2020.115116
Schaefer, T. J., Panda, P. K., & Wolford, R. W. (2025). Dengue Fever. In StatPearls. StatPearls Publishing. http://www.ncbi.nlm.nih.gov/books/NBK430732/
Selvarajah, D., Naing, C., Htet, N. H., & Mak, J. W. (2020). Loop-mediated isothermal amplification (LAMP) test for diagnosis of uncomplicated malaria in endemic areas: A meta-analysis of diagnostic test accuracy. Malaria Journal, 19(1), 211. https://doi.org/10.1186/s12936-020-03283-9
Shirshikov, F. V., & Bespyatykh, J. A. (2022). Loop-Mediated Isothermal Amplification: From Theory to Practice. Russian Journal of Bioorganic Chemistry, 48(6), 1159–1174. https://doi.org/10.1134/S106816202206022X
Shoushtari, M., Salehi-Vaziri, M., Roohvand, F., Arashkia, A., Jalali, T., & Azadmanesh, K. (2021). Taguchi array optimization of the reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for sensitive and rapid detection of dengue virus serotype 2. Biotechnology Letters, 43(11), 2149–2160. https://doi.org/10.1007/s10529-021-03175-1
Sinha, S., Singh, K., Ravi Kumar, Y. S., Roy, R., Phadnis, S., Meena, V., Bhattacharyya, S., & Verma, B. (2024). Dengue virus pathogenesis and host molecular machineries. Journal of Biomedical Science, 31(1), 43. https://doi.org/10.1186/s12929-024-01030-9
Soroka, M., Wasowicz, B., & Rymaszewska, A. (2021). Loop-Mediated Isothermal Amplification (LAMP): The Better Sibling of PCR? Cells, 10(8). https://doi.org/10.3390/cells10081931
Su, W., Jiang, L., Lu, W., Xie, H., Cao, Y., Di, B., Li, Y., Nie, K., Wang, H., Zhang, Z., & Xu, S. (2022). A Serotype-Specific and Multiplex PCR Method for Whole-Genome Sequencing of Dengue Virus Directly from Clinical Samples. Microbiology Spectrum, 10(5). https://doi.org/10.1128/spectrum.01210-22
Thiese, M. S. (2014). Observational and interventional study design types; an overview. Biochemia Medica, 24(2), 199–210. https://doi.org/10.11613/BM.2014.022
Vairaperumal, T., Lee, P.-T., & Liu, P.-Y. (2025). Portable Point-of-Care Diagnosis Platforms and Emerging Predictive Biomarkers for Rapid Detection of Severe Dengue Viral Infection. ACS Sensors, 10(5), 3302–3316. https://doi.org/10.1021/acssensors.5c00263
van den Elsen, K., Alvin, C. B. L., Sheng, H. J., & Dahai, L. (2023). Flavivirus nonstructural proteins and replication complexes as antiviral drug targets. Current Opinion in Virology, 59. https://doi.org/10.1016/j.coviro.2023.101305
WHO. (2009). Laboratory Diagnosis and Diagnostic Tests. In Dengue: Guidelines for Diagnosis, Treatment, Prevention and Control: New Edition. World Health Organization. https://www.ncbi.nlm.nih.gov/books/NBK143156/
WHO. (2024). Dengue and severe dengue. https://www.who.int/news-room/fact-sheets/detail/dengue-and-severe-dengue
Yang, N., Zhang, H., Han, X., Liu, Z., & Lu, Y. (2024). Advancements and applications of loop-mediated isothermal amplification technology: A comprehensive overview. Frontiers in Microbiology, 15. https://doi.org/10.3389/fmicb.2024.1406632
Yaren, O., McCarter, J., Phadke, N., Bradley, K. M., Overton, B., Yang, Z., Ranade, S., Patil, K., Bangale, R., & Benner, S. A. (2021). Ultra-rapid detection of SARS-CoV-2 in public workspace environments. PloS One, 16(2). https://doi.org/10.1371/journal.pone.0240524
Downloads
Published
How to Cite
Issue
Section
Citation Check
License
Copyright (c) 2026 Syafiq Maulana, Fatahillah Tsabit Fatoni
This work is licensed under a Creative Commons Attribution 4.0 International License.
