Browsing by Author "Ateya, L."
Now showing 1 - 2 of 2
- Results Per Page
- Sort Options
Item Development and Validation of Rapid Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification for Detection of Rift Valley Fever Virus(WILEY, 2023-01-30) Wekesa, F.; Wamalwa, M.; Oduor, R.; Binepal, Y.; Ateya, L.; Okumu, N.; M’kwenda, A.; Masaba, C.; Mukhaye, E.; Kenyatta University ; Kenya Agricultural Livestock and Research OrganizationRift Valley fever virus (RVFV) is a high‐priority zoonotic pathogen with the ability to cause massive loss during its outbreak within a very short period of time. Lack of a highly sensitive, instant reading diagnostic method for RVFV, which is more suitable for on‐site testing, is a big gap that needs to be addressed. The aim of this study was to develop a novel one‐step reverse transcription loop‐mediated isothermal amplification (RT‐LAMP) method for the rapid detection of RVFV. To achieve this, the selected RVFV M segment nucleotide sequences were aligned using Multiple Sequence Comparison by Log‐Expectation (MUSCLE) software in MEGA11 version 11.0.11 program to identify conserved regions. A 211 pb sequence was identified and six different primers to amplify it were designed using NEB LAMP Primer design tool version 1.1.0. The specificity of the designed primers was tested using primer BLAST, and a primer set, specific to RVFV and able to form a loop, was selected. In this study, we developed a single‐tube test based on calorimetric RT‐LAMP that enabled the visual detection of RVFV within 30 minutes at 65°C. Diagnostic sensitivity and specificity of the newly developed kit were compared with RVFV qRT‐PCR, using total RNA samples extracted from 118 blood samples. The colorimetric RT‐LAMP assay had a sensitivity of 98.36% and a specificity of 96.49%. The developed RT‐LAMP was found to be tenfold more sensitive compared to the RVFV qRT‐PCR assay commonly used in the confirmatory diagnosis of RVFV.Item Evaluation of Protective Efficacy of Inactivated Thermostable Vaccine Against Nairobi Sheep Disease Virus(Journal of Innovative Science and Research Technology, 2023-05) Muriuki, N.P.; Ithinji, G.D.; Nyamache, A.K.; Ateya, L.; Binepal, Y.S.; Wasonga, C.; Lutomiah, J.; Kiraithe, M.M.; Kenyatta University ; Kenya Agricultural and Livestock Research Organization (KALRO) ; University of Nairobi ; Kenya Medical Research Institute ; Kenya Veterinary Vaccine Production Institute (KEVEVAPI)Sheep and goats (n= 4/group) were inoculated with thermos-stabilized inactivated Nairobi sheep disease virus vaccine. Four unvaccinated animals for each group were kept as control. Vaccinated groups were given a booster vaccine dose 21 days later. Immune response was monitored by neutralizing antibody titers were determined by micro-plaque reduction neutralization test and confirmed by immunofluorescence assay. Two doses of the inactivated vaccine stimulated a strong immune response in the vaccinated animals. The vaccinated and mock group were challenged with virulent 1473 strain of the Nairobi sheep disease virus. All animals developed fever and viremia with varying degrees between sheep and goats post challenge. Mock vaccinated sheep developed high viremia levels relative to the vaccinated group and developed severe disease. In contrast, mock vaccinated goats showed a slight temperature compared to vaccinated goats. After challenge, two control sheep died from the disease whereas the vaccinated sheep survived. Vaccinated sheep suffered mild to moderate clinical reactions with pyrexia. Formalin inactivated vaccine fully protected the animals against the lethal 1473 challenge virus.
