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  1. Fischer K, Pickering B, Diederich S
    Methods Mol Biol, 2023;2610:17-29.
    PMID: 36534278 DOI: 10.1007/978-1-0716-2895-9_2
    Nipah virus (NiV) is an emerging, zoonotic paramyxovirus that is among the most pathogenic of viruses in humans. During the first reported outbreak of NiV in Malaysia and Singapore in the late 1990s, pigs served as an intermediate host, which enabled the transmission to humans. Although subsequent outbreaks in Asia only reported direct bat-to-human and human-to-human transmission, pigs are still considered a potential source for viral dissemination in the epidemiology of the disease. Thus, serological assays such as Enzyme-linked immunosorbent assay (ELISA) or virus neutralization test (VNT) represent powerful tools to characterize the serum antibody responses in NiV-infected pigs as well as to perform seroepidemiological surveillance studies on the potential circulation of NiV or NiV-related viruses among pig populations worldwide. This chapter describes both methods in detail. Furthermore, we discuss some of the major pitfalls and indicate how to avoid them.
  2. Kasloff SB, Leung A, Pickering BS, Smith G, Moffat E, Collignon B, et al.
    Sci Rep, 2019 03 26;9(1):5230.
    PMID: 30914663 DOI: 10.1038/s41598-019-40476-y
    In 1998 an outbreak of fatal encephalitis among pig farm workers in Malaysia and Singapore led to the discovery of Nipah henipavirus (NiV), a novel paramyxovirus closely related to Hendra henipavirus with case fatality rates of nearly 40%. Following its initial emergence nearly annual outbreaks of NiV have occurred in Bangladesh with a different, NiV Bangladesh, genotype, where the role of pigs in its transmission remains unknown. The present study provides the first report on susceptibility of domestic pigs to NiV Bangladesh following experimental infection, characterizing acute and long-term phases of disease and pathogenesis. All pigs were successfully infected with NiV Bangladesh following oronasal inoculation, with viral shedding confirmed by a novel genotype-specific qRT-PCR in oral, nasal and rectal excretions and dissemination from the upper respiratory tract to the brain, lungs, and associated lymphatic tissues. Unlike previous NiV Malaysia findings in pigs, clinical signs were absent, viremia was undetectable throughout the study, and only low level neutralizing antibody titers were measured by 28/29 days post-NiV-B infection. Results obtained highlight the need for continued and enhanced NiV surveillance in pigs in endemic and at-risk regions, and raise questions regarding applicability of current serological assays to detect animals with previous NiV-B exposure.
  3. Zhu W, Pickering B, Smith G, Pinette M, Truong T, Babiuk S, et al.
    Front Vet Sci, 2023;10:1120367.
    PMID: 36816187 DOI: 10.3389/fvets.2023.1120367
    INTRODUCTION: Nipah virus (NiV) and Hendra virus (HeV), of the genus Henipavirus, family Paramyxoviridae, are classified as Risk Group 4 (RG4) pathogens that cause respiratory disease in pigs and acute/febrile encephalitis in humans with high mortality.

    METHODS: A competitive enzyme-linked immunosorbent assay (cELISA) using a monoclonal antibody (mAb) and recombinant NiV glycoprotein (G) was developed and laboratory evaluated using sera from experimental pigs, mini pigs and nonhuman primates. The test depends on competition between specific antibodies in positive sera and a virus-specific mAb for binding to NiV-G.

    RESULTS: Based on 1,199 negative and 71 NiV positive serum test results, the cutoff value was determined as 35% inhibition. The diagnostic sensitivity and specificity of the NiV cELISA was 98.58 and 99.92%, respectively. When testing sera from animals experimentally infected with NiV Malaysia, the cELISA detected antibodies from 14 days post-infection (dpi) and remained positive until the end of the experiment (28 dpi). Comparisons using the Kappa coefficient showed strong agreement (100%) between the cELISA and a plaque reduction neutralization test (PRNT).

    DISCUSSION: Because our cELISA is simpler, faster, and gives comparable or better results than PRNT, it would be an adequate screening test for suspect NiV and HeV cases, and it would also be useful for epidemiological surveillance of Henipavirus infections in different animal species without changing reagents.

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