Comparison of Extraction Methods for the Detection of Tick-Borne Encephalitis Virus RNA in Goat Raw Milk and Cream Cheese

Author:

Müller Irene,Althof Nadine,Hoffmann Bernd,Klaus Christine,Schilling-Loeffler Katja,Falkenhagen Alexander,Johne ReimarORCID

Abstract

AbstractInfection with the tick-borne encephalitis virus (TBEV) can cause meningitis, meningoencephalitis and myelitis in humans. TBEV is an enveloped RNA virus of the family Flaviviridae, which is mostly transmitted via tick bites. However, transmission by consumption of virus-contaminated goat raw milk and goat raw milk products has also been described. Only a few methods have been reported for the detection of TBEV in food so far. Here, we compare different virus extraction methods for goat raw milk and goat raw milk cream cheese and subsequent detection of TBEV-RNA by RT-qPCR. Langat virus (LGTV), a naturally attenuated TBEV strain, was used for artificial contamination experiments. Mengovirus and the human coronavirus 229E were compared to assess their suitability to serve as internal process controls. Out of three tested extraction protocols for raw milk, sample centrifugation followed by direct RNA extraction from the aqueous interphase yielded the best results, with a recovery rate (RR) of 31.8 ± 4.9% for LGTV and a detection limit of 6.7 × 103 LGTV genome copies/ml. Out of two methods for cream cheese, treatment of the samples with TRI Reagent® and chloroform prior to RNA extraction showed the best RR of 4.7 ± 1.6% for LGTV and a detection limit of 9.4 × 104 LGTV genome copies/g. RRs of Mengovirus and LGTV were similar for both methods; therefore, Mengovirus is suggested as internal process control virus. The developed methods may be useful for screening or surveillance studies, as well as in outbreak investigations.

Funder

Bundesinstitut für Risikobewertung

Bundesinstitut für Risikobewertung (BfR)

Publisher

Springer Science and Business Media LLC

Subject

Virology,Health, Toxicology and Mutagenesis,Food Science,Epidemiology

Reference35 articles.

1. Althof, N., Trojnar, E., Böhm, T., Burkhardt, S., Carl, A., Contzen, M., Kilwinski, J., Mergemeier, S., Moor, D., Mäde, D., & Johne, R. (2019). Interlaboratory validation of a method for hepatitis E virus RNA detection in meat and meat products. Food and Environmental Virology, 11(1), 1–8. https://doi.org/10.1007/s12560-018-9360-6

2. Balogh, Z., Egyed, L., Ferenczi, E., Bán, E., Szomor, K. N., Takács, M., & Berencsi, G. (2012). Experimental infection of goats with tick-borne encephalitis virus and the possibilities to prevent virus transmission by raw goat milk. Intervirology, 55(3), 194–200. https://doi.org/10.1159/000324023

3. Brockmann, S. O., Oehme, R., Buckenmaier, T., Beer, M., Jeffery-Smith, A., Spannenkrebs, M., Haag-Milz, S., Wagner-Wiening, C., Schlegel, C., Fritz, J., Zange, S., Bestehorn, M., Lindau, A., Hoffmann, D., Tiberi, S., Mackenstedt, U., & Dobler, G. (2018). A cluster of two human cases of tick-borne encephalitis (TBE) transmitted by unpasteurised goat milk and cheese in Germany, May 2016. Euro Surveillance, 23(15), 17–00336. https://doi.org/10.2807/1560-7917.ES.2018.23.15.17-00336

4. Buczek, A. M., Buczek, W., Buczek, A., & Wysokińska-Miszczuk, J. (2022). Food-borne transmission of tick-borne encephalitis virus-spread, consequences, and prophylaxis. International Journal of Environmental Research and Public Health, 19(3), 1812. https://doi.org/10.3390/ijerph19031812

5. Chitimia-Dobler, L., Lindau, A., Oehme, R., Bestehorn-Willmann, M., Antwerpen, M., Drehmann, M., Hierl, T., Mackenstedt, U., & Dobler, G. (2021). Tick-borne encephalitis vaccination protects from alimentary tbe infection: Results from an alimentary outbreak. Microorganisms, 9(5), 889. https://doi.org/10.3390/microorganisms9050889

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