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  • 1
    Online Resource
    Online Resource
    Development of a Real-Time PCR Method for the Detection of European and Siberian Subtypes of Tick-Borne Encephalitis Virus
    MDPI AG ; 2023
    In:  Microbiology Research Vol. 14, No. 4 ( 2023-09-29), p. 1545-1558
    Details
    In: Microbiology Research, MDPI AG, Vol. 14, No. 4 ( 2023-09-29), p. 1545-1558
    Abstract: The tick-borne encephalitis virus (TBEV) is transmitted to humans through tick bites. In recent years, the appearance of the Siberian subtype of TBEV in Ixodes ricinus in Finland, together with deaths from the normally mild European subtype in the same country, have raised concerns about a possible spread of virulent variants of TBEV in Western Europe. Thus, there is a need to monitor the spread of strains, particularly of the European and Siberian subtypes. In this study, we develop a new real-time PCR method targeting Siberian and European subtypes of TBEV. The primers amplify a 176 bp fragment of the E gene, which is suitable for subsequent strain identification by Sanger sequencing. This study pioneers a new approach to primer design where the melting temperature (Tm) of primers annealed to representative mismatched target sequences is empirically determined and used to guide improvements in primer sequence. This allowed the range of TBEV strains detected to be extended to cover most European and Siberian strains tested, in addition to a strain of the Far-Eastern subtype. The limit of detection was 10–100 DNA copies per reaction and amplification efficiency varied between 83% and 94%, depending on the TBEV strain. Experimental determination of primer Tm proved to be a fruitful approach and will be a useful tool for future primer design and diagnostics.
    Type of Medium: Online Resource
    ISSN: 2036-7481
    URL: Article
    DOI: 10.3390/microbiolres14040106
    Language: English
    Publisher: MDPI AG
    Publication Date: 2023
    detail.hit.zdb_id: 2571057-6
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  • 2
    Online Resource
    Online Resource
    The Molecular Epidemiology of Enterovirus in a Birth Cohort in Nepal
    MDPI AG ; 2023
    In:  Microbiology Research Vol. 14, No. 3 ( 2023-07-20), p. 909-917
    Details
    In: Microbiology Research, MDPI AG, Vol. 14, No. 3 ( 2023-07-20), p. 909-917
    Abstract: Acute gastroenteritis (AGE) has a major impact on morbidity and mortality worldwide. The viral aetiology of diarrhoeal diseases may remain unknown due to limited diagnostic facilities. Non-polio enteroviruses (NPEVs) are the third most frequent pathogen detected in stool specimens from AGE cases, yet their potential role in AGE is uncertain. In Nepal, limited data are available on NPEVs, due to both the lack of an adequate surveillance program and the availability of tests. The global polio eradication initiative effort of the WHO has eradicated the incidence of poliomyelitis and acute flaccid paralysis (AFP) from many parts of the world, including Nepal. However, cases of AFP associated with NPEVs have been reported in different countries, including the neighbouring India. This study aims to investigate the diarrhoeal stool samples from a birth cohort until the age of 36 months for NPEVs and the genotype diversity of NPEV in community children with diarrhoea. A total of 280 longitudinal diarrhoeal stool samples that were negative for other enteric pathogens were tested using RT-PCRs. NPEVs was detected in 97 stool specimens (34.6%) and were significantly more frequent in infants up to one year of age. This study identified 17 various NPEV types, with the dominating species being Enterovirus B (EV-B). Ten different types of echoviruses were recorded in this study, with the two rare NPEVs B74 and A120. Based on prevalence, seasonality, and diversity, further studies are warranted to investigate the role of enterovirus in diarrhoeal disease.
    Type of Medium: Online Resource
    ISSN: 2036-7481
    URL: Article
    DOI: 10.3390/microbiolres14030063
    Language: English
    Publisher: MDPI AG
    Publication Date: 2023
    detail.hit.zdb_id: 2571057-6
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  • 3
    Online Resource
    Online Resource
    Transient Expression of Flavivirus Structural Proteins in Nicotiana benthamiana
    MDPI AG ; 2022
    In:  Vaccines Vol. 10, No. 10 ( 2022-10-06), p. 1667-
    Details
    In: Vaccines, MDPI AG, Vol. 10, No. 10 ( 2022-10-06), p. 1667-
    Abstract: Flaviviruses are a threat to public health and can cause major disease outbreaks. Tick-borne encephalitis (TBE) is caused by a flavivirus, and it is one of the most important causes of viral encephalitis in Europe and is on the rise in Sweden. As there is no antiviral treatment available, vaccination remains the best protective measure against TBE. Currently available TBE vaccines are based on formalin-inactivated virus produced in cell culture. These vaccines must be delivered by intramuscular injection, have a burdensome immunization schedule, and may exhibit vaccine failure in certain populations. This project aimed to develop an edible TBE vaccine to trigger a stronger immune response through oral delivery of viral antigens to mucosal surfaces. We demonstrated successful expression and post-translational processing of flavivirus structural proteins which then self-assembled to form virus-like particles in Nicotiana benthamiana. We performed oral toxicity tests in mice using various plant species as potential bioreactors and evaluated the immunogenicity of the resulting edible vaccine candidate. Mice immunized with the edible vaccine candidate did not survive challenge with TBE virus. Interestingly, immunization of female mice with a commercial TBE vaccine can protect their offspring against TBE virus infection.
    Type of Medium: Online Resource
    ISSN: 2076-393X
    URL: Article
    DOI: 10.3390/vaccines10101667
    Language: English
    Publisher: MDPI AG
    Publication Date: 2022
    detail.hit.zdb_id: 2703319-3
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