Development of an Antigen Capture Immunoassay Based on Monoclonal Antibodies Specific for Dengue Virus Serotype 2 Nonstructural Protein 1 for Early and Rapid Identification of Dengue Virus Serotype 2 Infections

Qiu, Li-wen; Di, Biao; Wen, Kai; Wang, Xin-shuai; Liang, Wenjie; Wang, Yadi; Pan, Yuxian; Wang, Ming; Ding, Yan‐Qing; Che, Xin

doi:10.1128/cvi.00212-08

Cited by 33 publications

(48 citation statements)

References 31 publications

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“…Preparation and identification of MAbs against NS1 of DENV1, DENV2, DENV3, and DENV4 were described in our previous studies (4,12,20). The characteristics of MAbs against NS1 of DENV1 and DENV2 were described in previous publications (4,12), and those of MAbs against NS1 of DENV3 and DENV4 are shown in Tables S1 and S2 in the supplemental material. The purified MAbs from ascitic fluids were conjugated with horseradish peroxidase (HRP; Sigma-Aldrich) by the periodate method (17).…”

Section: Serum Samples and Virusesmentioning

confidence: 99%

“…Another 30 acute-phase serum samples from DENV2-infected patients were collected in Guangdong Province, China, in 2001, as described in our previous report (12). Seven acute-phase serum samples from DENV3-infected patients were collected in Guangzhou in 2009.…”

Section: Serum Samples and Virusesmentioning

confidence: 99%

“…The production of epitope-specific monoclonal antibodies (MAbs) holds potential for the development of either group-specific or serotype-specific NS1 antigen assays. In previous studies, we developed DENV1-and DENV2-specific NS1 capture ELISAs using MAbs against serotype-specific NS1 (12,20). In the present study, the DENV3-specific and DENV4-specific NS1 capture ELISAs were further developed using well-characterized MAbs that recognized epitopes specific for DENV3 and DENV4 NS1.…”

mentioning

confidence: 99%

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Full Serotype- and Group-Specific NS1 Capture Enzyme-Linked Immunosorbent Assay for Rapid Differential Diagnosis of Dengue Virus Infection

Ding

Chen

et al. 2011

Clin Vaccine Immunol

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Dengue virus (DENV), a member of the Flavivirus family, has four distinct serotypes (DENV serotype 1 [DENV1], DENV2, DENV3, and DENV4) that require differentiation for the effective prevention of morbid disease. Early and rapid differentiation between flaviviruses remains challenging. Full assays combining four individual, serotype-specific and one group-specific nonstructural protein 1 (NS1) antigen capture enzymelinked immunosorbent assays (ELISAs) based on monoclonal antibodies (MAbs) against DENV NS1 were developed and validated. The sensitivities and specificities of the full NS1 ELISAs were evaluated with viral cultures and dengue acute-phase sera. Four serotype-specific NS1 ELISAs displayed high specificities for the detection and differentiation of appropriate serotypes. The group-specific NS1 ELISA was broadly reactive with the four dengue virus serotypes. None of the NS1 ELISAs displayed cross-reactivity with the other flaviviruses or samples from febrile patients with non-dengue virus infections. The full serotype-and group-specific MAb-based NS1 capture ELISAs may provide tools for the early detection and typing of dengue infection, which is preferable to reverse transcriptase PCR (RT-PCR) for the rapid differential diagnosis of dengue virus infection in the field.Dengue virus (DENV), a member of the Flavivirus family, has four distinct serotypes (DENV serotype 1 [DENV1], DENV2, DENV3, and DENV4). Infection with any of the four serotypes can cause disease ranging from dengue fever (DF) to dengue hemorrhagic fever (DHF)/dengue shock syndrome (DSS) (8). Subsequent heterologous infections may increase the risk of developing DHF or DSS (11). Patients with DHF or DSS can experience a sudden onset of hemorrhage or shock and may even die without appropriate management. Early diagnosis and management can reduce the morbidity and mortality of DHF or DSS (5). However, symptoms of dengue fever are not sufficiently specific for the accurate clinical differentiation of dengue from other febrile illnesses and hemorrhagic diseases, especially in areas where multiple tropical diseases, such as yellow fever, West Nile disease, Japanese encephalitis, and St. Louis encephalitis, are endemic. In addition, case identification has become more important in order to determine the correlations of different serotypes with disease severity (15). IgM antibody capture enzyme-linked immunosorbent assay (MAC-ELISA) has commonly been used for routine diagnosis. Anti-dengue virus IgM antibody detection, however, is more challenging because dengue virus antibodies cross-react with other flaviviruses (10). Viral RNA detection assays, such as one-step TaqMan real-time reverse transcriptase PCR (RT-PCR), provide a promising sensitivity rate and rapid diagnosis at the acute stage. However, the molecular approach is costly because it requires specialized laboratory equipment and experienced technicians (14). These represent notable limitations in many developing countries where dengue disease is endemic (14). Nonstructural protein 1 (NS1)...

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Section: Serum Samples and Virusesmentioning

confidence: 99%

Section: Serum Samples and Virusesmentioning

confidence: 99%

mentioning

confidence: 99%

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Full Serotype- and Group-Specific NS1 Capture Enzyme-Linked Immunosorbent Assay for Rapid Differential Diagnosis of Dengue Virus Infection

Ding

Chen

et al. 2011

Clin Vaccine Immunol

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“…The slides were incubated with the monoclonal antibodies (MAbs) against Mp1p at a concentration of 10 g/ml in a humid chamber for 60 min at 37°C. The MAb was prepared by a procedure described previously (44,45). Briefly, BALB/c mice were immunized intradermally with 50 mg of purified Mp1p emulsified with Freund's adjuvant at 10-day intervals for 8 weeks followed by intravenously administered boosters for 3 days before fusion.…”

Section: P Marneffei Blastomyces Dermatitidis Histoplasma Capsulatmentioning

confidence: 99%

Unraveling the Molecular Basis of Temperature-Dependent Genetic Regulation in Penicillium marneffei

et al. 2013

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c Penicillium marneffei is an opportunistic fungal pathogen endemic in Southeast Asia, causing lethal systemic infections in immunocompromised patients. P. marneffei grows in a mycelial form at the ambient temperature of 25°C and transitions to a yeast form at 37°C. The ability to alternate between the mycelial and yeast forms at different temperatures, namely, thermal dimorphism, has long been considered critical for the pathogenicity of P. marneffei, yet the underlying genetic mechanisms remain elusive. Here we employed high-throughput sequencing to unravel global transcriptional profiles of P. marneffei PM1 grown at 25 and 37°C. Among ϳ11,000 protein-coding genes, 1,447 were overexpressed and 1,414 were underexpressed at 37°C. Counterintuitively, heat-responsive genes, predicted in P. marneffei through sequence comparison, did not tend to be overexpressed at 37°C. These results suggest that P. marneffei may take a distinct strategy of genetic regulation at the elevated temperature; the current knowledge concerning fungal heat response, based on studies of model fungal organisms, may not be applicable to P. marneffei. Our results further showed that the tandem repeat sequences (TRSs) are overrepresented in coding regions of P. marneffei genes, and TRS-containing genes tend to be overexpressed at 37°C. Furthermore, genomic sequences and expression data were integrated to characterize gene clusters, multigene families, and species-specific genes of P. marneffei. In sum, we present an integrated analysis and a comprehensive resource toward a better understanding of temperature-dependent genetic regulation in P. marneffei.

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“…The preparation of MAb against nucleoprotein of influenza virus A H1N1 WSN strain was performed as previously described, with some modifications (30,50). Briefly, BALB/c mice were first immunized intraperitoneally with 10 g of recombinant nucleoprotein of influenza A virus, and were then given five boosters of the same doses of nucleoprotein of influenza A virus at 10-day intervals.…”

Section: Participantsmentioning

confidence: 99%

Differences in Antibody Responses of Individuals with Natural Infection and Those Vaccinated against Pandemic H1N1 2009 Influenza

Chan

Hung

et al. 2011

Clin. Vaccine Immunol.

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The differential antibody response measured by the commonly used hemagglutination inhibition (HI) and microneutralization (MN) assays in patients with natural infection and vaccination has not been fully assessed. HI and conventional MN (CMN) assays were performed on sera from 651 patients with natural infection by pandemic H1N1 2009 influenza virus and on sera from 567 recipients of the corresponding vaccine. Surprisingly, the overall seroprotection rates determined by CMN and HI assays in vaccine recipients were only 44.8 and 35.1%, respectively. Antibody titers measured by the CMN assay was significantly higher than that obtained by HI assay in vaccine recipients aged >50 years, but these titers were not significantly different among younger vaccine recipients. In contrast, the HI titer was greater than the CMN titer for the age group from 16 to 29 years but was not significantly different in other age groups for natural infection. Lower antibody levels were found in both naturally infected patients and immunized recipients in the older than in the younger age groups, but naturally infected patients exhibited higher HI and CMN titers than did the corresponding vaccine recipients. In addition, we developed a rapid fluorescent focus microneutralization (FFMN) assay to test sera from naturally infected patients. The FFMN assay has a better correlation with CMN than with HI ( ‫؍‬ 0.810 versus 0.684), which is expected of neutralizing antibody mainly targeted toward the inhibition of viral entry into cells. The higher antibody level elicited by natural infection than by vaccination may be related to differences between antigen presentation by the intramuscular route of vaccination and mucosal viral replication in mucosal cells of the respiratory tract.

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Development of an Antigen Capture Immunoassay Based on Monoclonal Antibodies Specific for Dengue Virus Serotype 2 Nonstructural Protein 1 for Early and Rapid Identification of Dengue Virus Serotype 2 Infections

Cited by 33 publications

References 31 publications

Full Serotype- and Group-Specific NS1 Capture Enzyme-Linked Immunosorbent Assay for Rapid Differential Diagnosis of Dengue Virus Infection

Full Serotype- and Group-Specific NS1 Capture Enzyme-Linked Immunosorbent Assay for Rapid Differential Diagnosis of Dengue Virus Infection

Unraveling the Molecular Basis of Temperature-Dependent Genetic Regulation in Penicillium marneffei

Differences in Antibody Responses of Individuals with Natural Infection and Those Vaccinated against Pandemic H1N1 2009 Influenza

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