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Войти в исследования Journal of virological methods · 2025
Development of a LAMP-based diagnostic method for HTLV-1 using Iranian clinical samples.
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Douzandegan Yousef, Kheirabad Ali Kargar, Mozhgani Sayed-Hamidreza, Tavoosidana Gholamreza, Foroushani Abbas Rahimi, Norouzi Mehdi
Аннотация
BACKGROUND: Human T-lymphotropic virus type 1 (HTLV-1) is a delta-retrovirus responsible for severe diseases such as adultT-cell leukemia/lymphoma (ATL) and HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). Accurate andaccessible diagnostic tools are required, especially in endemic regions such as Iran, to manage reasonably and prevent HTLV-1 infections.
OBJECTIVE: This study aimed to develop and validate an HTLV-1 detection protocol using loop-mediated isothermal amplification (LAMP) method on clinical samples from Iranianpatients.
METHODS: A LAMP assay was designed using optimized primers targeting the HTLV-1 genome. It was validated with clinical samples, and its sensitivity and specificity were evaluated compared to the polymerase chain reaction (PCR)method. Reaction conditions were optimized for fast and specific amplification, and results were visually analyzed utilizing a Luminator device and a real-time thermocycler.
RESULTS: The LAMP assay revealed a sensitivityof 100 % and specificity of 94.7 % for HTLV-1 detection. Within 15-30 min, the method produced results in isothermal conditionswith good performance. The fluorescence-based detection approach is easy touse. It does not need a sophisticated laboratory facility and has a limit of detection of 2.5 copies/µL.
CONCLUSION: This study demonstrates the feasibility of a LAMP-based method for HTLV-1 detection that is rapid, cost-effective, and accessible compared to PCR. For this reason,this alternative could enhance HTLV-1 diagnosis in both qualitative or quantitative form in endemic and low-resource scenarios. Future research should explore further validation in more diverse populations and integration with newtechnologies for digital detection.
OBJECTIVE: This study aimed to develop and validate an HTLV-1 detection protocol using loop-mediated isothermal amplification (LAMP) method on clinical samples from Iranianpatients.
METHODS: A LAMP assay was designed using optimized primers targeting the HTLV-1 genome. It was validated with clinical samples, and its sensitivity and specificity were evaluated compared to the polymerase chain reaction (PCR)method. Reaction conditions were optimized for fast and specific amplification, and results were visually analyzed utilizing a Luminator device and a real-time thermocycler.
RESULTS: The LAMP assay revealed a sensitivityof 100 % and specificity of 94.7 % for HTLV-1 detection. Within 15-30 min, the method produced results in isothermal conditionswith good performance. The fluorescence-based detection approach is easy touse. It does not need a sophisticated laboratory facility and has a limit of detection of 2.5 copies/µL.
CONCLUSION: This study demonstrates the feasibility of a LAMP-based method for HTLV-1 detection that is rapid, cost-effective, and accessible compared to PCR. For this reason,this alternative could enhance HTLV-1 diagnosis in both qualitative or quantitative form in endemic and low-resource scenarios. Future research should explore further validation in more diverse populations and integration with newtechnologies for digital detection.