Indirect ELISA and Indirect Immunofluorescent Antibody Assay for Detecting the Antibody against Murine Norovirus S7 in Mice

Kitagawa, Yota; Tohya, Yukinobu; Ike, Fumio; Kajita, Ayako; Park, Sang-Jin; Ishii, Yoshiyuki; Kyuwa, Shigeru; Yoshikawa, Yasuhiro

doi:10.1538/expanim.59.47

Cited by 30 publications

(30 citation statements)

References 20 publications

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“…Those constructs produce viral progeny, but the infectivity of the human strains could not be demonstrated because of the lack of a permissive cell system. In contrast, a construct of an MNV-S7 strain, which was initially isolated from mouse stool in Japan (15), produced infectious virions in several cell lines, including RAW264.7 cells and other non-mouse-derived cell lines such as HEK293T cells and COS7 cells. Taken together with the results of Guix et al (10), our results showed that, if the norovirus genome is provided intracellularly, infectious particles can be produced even in cells that are not…”

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confidence: 99%

Functional receptor molecules CD300lf and CD300ld within the CD300 family enable murine noroviruses to infect cells

Haga

Fujimoto

Takai‐Todaka

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

160

154

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Norovirus is the leading cause of acute gastroenteritis worldwide. Since the discovery of human norovirus (HuNoV), an efficient and reproducible norovirus replication system has not been established in cultured cells. Although limited amounts of virus particles can be produced when the HuNoV genome is directly transfected into cells, the HuNoV cycle of infection has not been successfully reproduced in any currently available cell-culture system. Those results imply that the identification of a functional cell-surface receptor for norovirus might be the key to establishing a norovirus culture system. Using a genome-wide CRISPR/Cas9 guide RNA library, we identified murine CD300lf and CD300ld as functional receptors for murine norovirus (MNV). The treatment of susceptible cells with polyclonal antibody against CD300lf significantly reduced the production of viral progeny. Additionally, ectopic CD300lf expression in nonsusceptible cell lines derived from other animal species enabled MNV infection and progeny production, suggesting that CD300lf has potential for dictating MNV host tropism. Furthermore, CD300ld, which has an amino acid sequence in the N-terminal region of its extracellular domain that is highly homologous to that of CD300lf, also functions as a receptor for MNV. Our results indicate that direct interaction of MNV with two cell-surface molecules, CD300lf and CD300ld, dictates permissive noroviral infection.

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mentioning

confidence: 99%

Functional receptor molecules CD300lf and CD300ld within the CD300 family enable murine noroviruses to infect cells

Haga

Fujimoto

Takai‐Todaka

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

160

154

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“…As these previous studies indicate that the effects of MNV infection on experiments in mice cannot be generalized, MNV-free mice are recommended for use in biomedical research. Thus, MNV infection in laboratory mouse colonies are monitored periodically by immunological methods such as the enzyme-linked immunosorbent assay [9] and multiplexed fluorescent immunoassay [8], because MNV is believed to comprise a single serogroup [16]. MNV infection in colonies may also be monitored by conventional reverse transcription-polymerase chain reaction (RT-PCR) assays including nested RT-PCR [8], [26]–[30].…”

Section: Introductionmentioning

confidence: 99%

A Broadly Reactive One-Step SYBR Green I Real-Time RT-PCR Assay for Rapid Detection of Murine Norovirus

et al. 2014

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A one-step SYBR Green I real-time RT-PCR assay was developed for the detection and quantification of a broad range of murine noroviruses (MNVs). The primer design was based on the multiple sequence alignments of 101 sequences of the open reading frame (ORF)1−ORF2 junction of MNV. The broad reactivity and quantitative capacity of the assay were validated using 7 MNV plasmids. The assay was completed within 1 h, and the reliable detection limit was 10 copies of MNV plasmid or 0.063 median tissue culture infective doses per milliliter of RAW264 cell culture-propagated viruses. The diagnostic performance of the assay was evaluated using 158 mouse fecal samples, 91 of which were confirmed to be positive. The melting curve analysis demonstrated the diversity of MNV in the samples. This is the first report of a broadly reactive one-step SYBR Green I real-time RT-PCR assay for detecting of MNVs. The rapid and sensitive performance of this assay makes it a powerful tool for diagnostic applications.

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“…This virus is highly prevalent among laboratory mice, and it is symptomless in immunologically normal mice [3,9,10,14,17,18,23]. Karst et al showed that susceptibility to this virus is not substantially increased by deleting the recombination activating gene 1 (Rag1) or Rag2 gene-dependent specific immunity [7].…”

Section: Introductionmentioning

confidence: 99%

“…serological [5,6,9,18] and rT-PCr assays [2,5,6,10,14] have been widely used to detect MnV infections. in particular, the use of rT-PCr has shown that mice experimentally inoculated with an MnV strain (MnV-2, MnV-3, or MnV-4) have detectable viruses in the feces from day 2 to week 8 post infection [5], revealing that mice can become persistently infected and shed virus in the feces for prolonged periods.…”

Section: Introductionmentioning

confidence: 99%

A Pitfall in Mouse Norovirus (MNV) Detection in Fecal Samples Using RT-PCR, and Construction of New MNV-Specific Primers

et al. 2013

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Abstract:The murine norovirus (MNV), which belongs to the Caliciviridae family, is prevalent in laboratory mice. Since this virus affects macrophages and dendritic cells, infected mice are not suitable for immunological investigations, making it important to detect MNV infections accurately. When we tested RNA extracts derived from mouse feces for MNV detection using nested RT-PCR with a set of MNV-specific primers reported by Goto et al. (Exp. Anim. 58: 135-140, 2009), we found that these primers amplified not only an MNV-specific signal but also amplified a relatively weak signal with a size almost identical to that of the specific signal. Analysis of the nucleotide sequence of this amplified signal revealed that it was at least 98% identical to the exophosphatase gene of a commensal bacterium, Bacteroides vulgatus. Subsequent analysis showed that the signal amplified with a pair of nested primers was from DNA derived from B. vulgatus, which is sometimes present in SPF laboratory mouse feces, and the nested primers used were both partly homologous with the B. vulgatus nucleotide sequence. We thus designed a new set of nested RT-PCR primers that was not cross-reactive with the B. vulgatus genome. PCR products amplified by the newly designed primers were at least 89.3% identical to the MNV RNA polymerase gene in all cases. Our findings demonstrated that the primer set we designed was suitable for detecting an MNV-specific signal without crossreacting with B. vulgatus DNA in mouse feces.

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Indirect ELISA and Indirect Immunofluorescent Antibody Assay for Detecting the Antibody against Murine Norovirus S7 in Mice

Cited by 30 publications

References 20 publications

Functional receptor molecules CD300lf and CD300ld within the CD300 family enable murine noroviruses to infect cells

Functional receptor molecules CD300lf and CD300ld within the CD300 family enable murine noroviruses to infect cells

A Broadly Reactive One-Step SYBR Green I Real-Time RT-PCR Assay for Rapid Detection of Murine Norovirus

A Pitfall in Mouse Norovirus (MNV) Detection in Fecal Samples Using RT-PCR, and Construction of New MNV-Specific Primers

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