Isolation and Identification of Duck Tembusu Virus Strain LH and Development of Latex-Agglutination Diagnostic Method for Rapid Detection of Antibodies

Wang, Quanxi; Wen, Yaping; Huang, Yifan; Wu, Yijian; Cai, Yilong; Xu, Lihui; Wang, Changkang; Li, Ang; Wu, Benhong; Chen, Ji‐Long

doi:10.1637/10795-021114-reg

Cited by 8 publications

(4 citation statements)

References 16 publications

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“…While sexual transmission has not been documented for other flavivirus infections, some members of this family have been recovered from fecal samples of avian hosts. The RNA of Tembusu virus was detected in duck feces and cloacal swabs 30 , 31 . Infectious particles of West Nile virus (WNV) were isolated from oral and cloacal swab samples of dead birds 32 .…”

Section: Discussionmentioning

confidence: 99%

Zika virus shedding in the stool and infection through the anorectal mucosa in mice

Deng

et al. 2018

Emerging Microbes & Infections

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Zika virus (ZIKV) has elicited global concern due to its unique biological features, unusual transmission routes, and unexpected clinical outcomes. Although ZIKV transmission through anal intercourse has been reported in humans, it remains unclear if ZIKV is detectable in the stool, if it can infect the host through the anal canal mucosa, and what the pathogenesis of such a route of infection might be in the mouse model. Herein, we demonstrate that ZIKV RNA can be recovered from stools in multiple mouse models, as well as from the stool of a ZIKV patient. Remarkably, intra-anal (i.a.) inoculation with ZIKV leads to efficient infection in both Ifnar1−/− and immunocompetent mice, characterized by extensive viral replication in the blood and multiple organs, including the brain, small intestine, testes, and rectum, as well as robust humoral and innate immune responses. Moreover, i.a. inoculation of ZIKV in pregnant mice resulted in transplacental infection and delayed fetal development. Overall, our results identify the anorectal mucosa as a potential site of ZIKV infection in mice, reveal the associated pathogenesis of i.a. infection, and highlight the complexity of ZIKV transmission through anal intercourse.

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Section: Discussionmentioning

confidence: 99%

Zika virus shedding in the stool and infection through the anorectal mucosa in mice

Deng

et al. 2018

Emerging Microbes & Infections

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“…In general, TMUV is isolated using embryonated duck/chicken eggs416, primary duck/chicken embryo cell cultures217, mammalian cell lines, and mosquito cell lines18 for initial virology diagnosis purposes. Serological methods of virus neutralization19, ELISA202122, and fluorescent antibody23 tests are also commonly used for the detection of TMUV antigens or specific antibodies.…”

mentioning

confidence: 99%

Advanced uracil DNA glycosylase-supplemented real-time reverse transcription loop-mediated isothermal amplification (UDG-rRT-LAMP) method for universal and specific detection of Tembusu virus

Tang

Chen

Diao

2016

Sci Rep

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Tembusu virus (TMUV) is a mosquito-borne flavivirus which threatens both poultry production and public health. In this study we developed a complete open reading frame alignment-based rRT-LAMP method for the universal detection of TUMV. To prevent false-positive results, the reaction was supplemented with uracil DNA glycosylase (UDG) to eliminate carryover contamination. The detection limit of the newly developed UDG-rRT-LAMP for TMUV was as low as 100 copies/reaction of viral RNA and 1 × 100.89 − 1 × 101.55 tissue culture infectious dose/100 μL of viruses. There were no cross-reactions with other viruses, and the reproducibility of the assay was confirmed by intra- and inter-assay tests with variability ranging from 0.22–3.33%. The new UDG-rRT-LAMP method for TMUV produced the same results as viral isolation combined with RT-PCR as the “gold standard” in 96.88% of cases for 81 clinical samples from subjects with suspected TMUV infection. The addition of UDG can eliminate as much as 1 × 10−16 g/reaction of contaminants, which can significantly reduce the likelihood of false-positive results during the rRT-LAMP reaction. Our result indicated that our UDG-rRT-LAMP is a rapid, sensitive, specific, and reliable method that can effectively prevent carryover contamination in the detection of TMUV.

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“…TMUV was associated with the explosive epizootics that occurred in commercial duck flocks in China in 2010 and in Southeast Asian countries later on [ 4 , 5 , 6 ]. The virus is transmitted among susceptible duck flocks by multiple routes (e.g., mosquito bites, infectious aerosol inhalation and eating contaminated feed) and results in severe egg drop and fatal encephalitis [ 7 , 8 , 9 ]. Because of the widespread duck farms and diversified duck housing systems in China and Southeastern Asia, vaccination is the preferred prophylactic countermeasure against highly contagious TMUV disease.…”

Section: Introductionmentioning

confidence: 99%

Amino Acid Substitutions in NS5 Contribute Differentially to Tembusu Virus Attenuation in Ducklings and Cell Cultures

Sun

Zhang

et al. 2021

Viruses

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Tembusu virus (TMUV), a highly infectious pathogenic flavivirus, causes severe egg-drop and encephalitis in domestic waterfowl, while the determinants responsible for viral pathogenicity are largely unknown. In our previous studies, virulent strain JXSP2-4 had been completely attenuated by successive passages in BHK-21 cells and the avirulent strain was designated as JXSP-310. Based on the backbone of JXSP2-4, a series of chimeric viruses were generated according to the amino acid substitutions in NS5 and their infectivities were also analyzed in cell cultures and ducklings. The results showed that the viral titers of RNA-dependent RNA polymerase (RdRp) domain-swapped cheimeric mutant (JXSP-310RdRp) in cells and ducklings were both markedly decreased compared with JXSP2-4, indicating that mutations in the RdRp domain affected viral replication. There are R543K and V711A two amino acid substitutions in the RdRp domain. Further site-directed mutagenesis showed that single-point R543K mutant (JXSP-R543K) exhibited similar replication efficacy compared with JXSP2-4 in cells, but the viral loads in JXSP-R543K-infected ducklings were significantly lower than that of JXSP2-4 and higher than JXSP-310RdRp. Surprisingly, the single-point V711A mutation we introduced rapidly reverted. In addition, qRT-PCR and Western blot confirmed that the mutations in the RdRp domain significantly affected the replication of the virus. Taken together, these results show that R543K substitution in the RdRp domain impairs the in vivo growth of TMUV, but sustaining its attenuated infectivity requires the concurrent presence of the V711A mutation.

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Isolation and Identification of Duck Tembusu Virus Strain LH and Development of Latex-Agglutination Diagnostic Method for Rapid Detection of Antibodies

Cited by 8 publications

References 16 publications

Zika virus shedding in the stool and infection through the anorectal mucosa in mice

Zika virus shedding in the stool and infection through the anorectal mucosa in mice

Advanced uracil DNA glycosylase-supplemented real-time reverse transcription loop-mediated isothermal amplification (UDG-rRT-LAMP) method for universal and specific detection of Tembusu virus

Amino Acid Substitutions in NS5 Contribute Differentially to Tembusu Virus Attenuation in Ducklings and Cell Cultures

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