Anti‐norovirus polyclonal antibody and its potential for development of an antigen‐ELISA

Okame, Michio; Shiota, Tomoyuki; Hansman, Grant S.; Takagi, Makiko; Yagyu, Fumihiro; Takanashi, Sayaka; Phan, Tung Gia; Shimizu, Yuko; Kohno, Hirotada; Okitsu, Shoko; Ushijima, Hiroshi

doi:10.1002/jmv.20906

Cited by 17 publications

(11 citation statements)

References 24 publications

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“…In comparison with other previous immunological methods such as ELISA and the immune-chromato- Table 3. graphic test (Khamrin et al, 2008;Takanashi et al, 2008), the detection limit of LAT (10 6 -10 7 genomic copies per gram of stool) was comparable. In our study, the specificity of the LAT for NoVs was 100%, which was comparable and even better that those from ELISA and the immune-chromatographic test (88-96%) (Rabenau et al, 2003;de Bruin et al, 2006;OkitsuNegishi et al, 2006;Okame et al, 2007;Khamrin et al, 2008;Takanashi et al, 2008). However, the sensitivity of LAT was only 35%, which was lower than that previous ELISA (31-90%) and the immune-chromatographic test (70-79%).…”

Section: Discussioncontrasting

confidence: 71%

Development of a latex agglutination test for norovirus detection

et al. 2010

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Norovirus (NoV) is the leading cause of acute gastroenteritis worldwide. Currently, reverse transcription polymerase chain reaction (RT-PCR) is used commonly to detect NoVs in both clinical and environmental samples. However, RT-PCR requires expensive equipment and cannot be performed on site. In this study, a latex agglutination test (LAT) using antibody-labeled latex beads for detecting NoVs was developed. Two kinds of polyclonal antibodies, one generated from synthetic peptides and the other from E. coli-expressed NoV capsid proteins, were used to develop the LAT. Each of these polyclonal antibodies was immobilized on the surface of latex beads and tested for the ability to detect NoVs. Under optimized conditions, our LAT detected GII.4 NoV at concentrations as low as 3.3x10(5) RT-PCR units/ml in stool samples. The detection limit for the LAT was approximately 1.7 103 RT-PCR units. Forty-eight stool samples were tested for NoVs using this LAT. In comparison with an RT-PCR assay, the sensitivity and specificity of the LAT were 35% and 100%, respectively. With further optimization, this LAT used with appropriate antibodies could be applied for convenient detection of NoVs in clinical diagnosis and food monitoring.

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Section: Discussioncontrasting

confidence: 71%

Development of a latex agglutination test for norovirus detection

et al. 2010

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“…Okame et al [33] developed a cross-reactive sandwich ELISA for detection of multiple GI and GII strains with a similar lower detection limit (0.0024 μg/mL) [33], but their assay required five antibodies (four primaries and one labeled secondary). Conversely, the capture ELISA presented in this study needed only one primary (for capture) and one labeled secondary antibody (for detection), drastically reducing assay time and reagent need.…”

Section: The "Signal-down" Capture Elisamentioning

confidence: 99%

“…Current approaches for detecting norovirus in clinical and environmental samples utilize a combination of electron microscopy techniques [20][21][22][23][24], molecular detection assays [7,10,17,[25][26][27] and immunological methods [25,[28][29][30][31][32][33]. Each approach strives for maximizing sensitivity and specificity while minimizing the time for detection [34].…”

Section: Introductionmentioning

confidence: 99%

Design and Evaluation of Three Immuno-based Assays for Rapid Detection of Human Norovirus Virus-like Particles

Broglie¹,

Moore²,

Jaykus³

et al. 2014

J Anal Bioanal Tech

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This study designed and evaluated three versatile immuno-based assays for the rapid detection of GI.1 norovirus virus-like particles at low (0-3.0 μg/mL) levels: 1) enzymatic absorbance-based ELISAs, 2) a fluorescentbased immunoassay, and 3) a "signal-down" capture ELISA. Variables including controllable variations in assay format (indirect or sandwich), assay time, binding sequence, and reporter molecule (fluorophore or enzyme) were thoroughly investigated and optimized in all three assays. Selectivity tests in the three-hour absorbance-based ELISA using VLPs representing two GI and two GII strains indicated the assays were selective to GI strains over GII strains. The three-hour enzymatic absorbance-based assay turned out to be a robust and rapid method capable of detecting GI.I VLPs in the range of 0.037 to 0.555 μg/mL, and the three-hour fluorescent immunoassay was capable of detecting VLPs in a high concentration range of 0.5-2.0 μg/mL under optimized conditions. The "signal-down" capture ELISA was conceptually demonstrated for the detection of VLPs at concentrations in excess of 1.0 μg/ mL, but did not appear to be suitable for quantifying VLPs under its current conditions. The methods reported here provide proof-of-concept that various ELISA-type approaches could be further developed to provide robust norovirus detection assays having various detection ranges, limits, and linearity.

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“…Recently, HuNoV strains belonging to GI and GII were subdivided into at least 15 and 18 genotypes, respectively (4,5). Of these, HuNoV GII/4 has been reported as the most prevalent genotype in causing acute (4,6).…”

Section: Human Norovirus (Hunov) Is In the Familymentioning

confidence: 99%