Development of LAMP assay for detection on lumpy skin disease virus in cattle
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Abstract
Lumpy skin disease (LSD), caused by a virus of the genus Capripoxvirus (CaPVs), represents a significant economic concern in cattle livestock management. LSD has the potential for rapid transmission, facilitated by blood-sucking insects that serve as vectors for the virus (LSDV). Additionally, transmission can occur through direct contact with infected animals, via the semen of an infected breeder, or transplacentally. The disease is characterized by fever, enlarged lymph nodes, firm and circumscribed nodules on the skin, and ulcerative lesions, particularly on the mucous membranes of the mouth. A key to preventing the spread of LSDV in cattle is rapid and accurate detection. Loop-mediated isothermal amplification (LAMP) assay is a novel technique that works by amplifying the specific target nucleic acid at a constant temperature, which is simple, fast, and easy to use in the field. The objective of this study was to develop a prototype LAMP assay to detect LSDV from various cattle samples and compare it to the conventional PCR assay. The results indicated that, of the 186 samples collected (comprising 100 EDTA blood samples and 86 skin scab samples) from cattle exhibiting clinical signs of lumpy skin disease (LSD), 56.98 % (106/186) tested positive for LSDV using the PCR assay. In contrast, 63.97 % (119/186) tested positive for LSDV using the LAMP assay, suggesting that the LAMP assay demonstrates higher sensitivity compared to the conventional PCR assay. Moreover, sensitivity analysis showed a detection limit of 10-5ng/µL. The specificity analysis test showed no cross-detection with other infectious agents. Additionally, the nucleotide alignment was 100 % identical to the LSDV strains from China, Hong Kong, and Vietnam. The LAMP assay detailed in this report is simple, cost-effective, highly sensitive, and particularly well-suited for the diagnosis of LSDV in laboratories with limited equipment and in resource-constrained rural settings.
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References
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