Location: Zoonotic and Emerging Disease Research
Project Number: 3022-32000-027-017-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Apr 1, 2024
End Date: Apr 1, 2026
Objective:
The objectives are as follows: 1) Develop and evaluate novel diagnostics for high consequence pathogens including CCHF, Nipah and Hendra virus; to perform surveillance of CCHF in the Czech Republic and neighboring areas as feasible; to evaluate a novel multiplex assay for the detection of antibodies to high risk pathogens in endemic countries.
Approach:
UCLA and NBAF scientists will partner with the Czech University of Life Sciences to develop protocols to collect Hyalomma sp. (all stages) from horses and Rhipicephalus sp. (all stages) from dogs in the Czech Republic, Romania and Bulgaria through the Citizen science programs and with the help of veterinarians and veterinary students (contacts already established in all three countries) in Years 1 and 2. The total DNA and RNA will be extracted from individual tick homogenates and used for tick identification by established protocols. Specifically, DNA will be used for amplification and sequencing of partial 16S rDNA and COI genes for tick identification and gltA and rompA genes for detection of R. conorii. RNA will be used for a nested RT-PCR targeting the S and M segment to detect CCHFv. Positive samples for either pathogen will be genotyped to better understand their epidemiology. Amplification and sequencing of the three spacers, dksA-xerC, mppA-purC and rpmE-tRNAfMet will be used to genotype R. conorii while segments S and M will be sequenced to genotype CCHFv. In addition, blood samples from horses and dogs will be collected and used for detection of antibodies (IgG/IgM) by commercial ELISA kits for CCHFv and R. conorii.
We will also partner Dr. Martine Peeters and Dr. Eric Delaporte in Montpellier, France to develop a multiplex assay to identify pathogens of interest, test banked and prospectively collected samples and train investigators in high-risk regions of the world in utilization of the assay. Samples may also be tested from existing sample banks and collected from France and/or other sites in Europe. These novel assays will be compared to commercially available assays.
