Method for the immunological diagnosis of a sample with a potential infection with an arbovirus and test kits suitable for this purpose
Inventors
Soutschek, Erwin • BOECHER, Oliver • NOELTING, Christina
Assignees
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Publication Number
US-10921323-B2
Publication Date
2021-02-16
Expiration Date
Abstract
The present invention relates to a method for the immunological diagnosis of a sample from a patient with a potential infection with an arbovirus, wherein a) a sample is brought into contact with a plurality of antigens that, separated from each other, are applied to a solid phase, wherein at least the following antigens are used: aa) the “non-structural protein 1” of a first arbovirus or an immunologically reactive part thereof having at least 8 amino acids,bb) an “envelope protein” of a first arbovirus or an immunologically reactive part thereof having at least 8 amino acids,cc) an “envelope protein” of a second arbovirus or an immunologically reactive part thereof having at least 8 amino acids,dd) the “non-structural protein 1” of a third arbovirus or an immunologically reactive part thereof having at least 8 amino acids andee) an “envelope protein” of a third arbovirus or an immunologically reactive part thereof having at least 8 amino acids, b) the solid phase is washed to separate non-specific bindings,c) the immune complex formed on the solid phase is converted into a signal andd) the test method is evaluated by comparing the relative signal strengths.
Core Innovation
The invention relates to an immunological differential diagnosis method for a sample from a human patient with a potential infection with an arbovirus. A sample is brought into contact with a plurality of antigens separated from each other and applied to a solid phase. The antigen panel includes non-structural protein 1 (NS1) and envelope protein antigens from Dengue virus serotype 2, envelope protein antigens from Chikungunya virus, and NS1 and envelope protein antigens from Zika virus, as immunologically reactive parts having at least 40 amino acids.
After contact, the solid phase is washed to separate non-specific bindings. Anti-human IgG and/or anti-human IgM antibodies coupled to a signal-generating agent are added, and any immune complex formed is detected by a signal from the signal-generating agent. The generated signal is compared to a negative and positive control, and the presence and/or absence of IgG and/or IgM antibodies against the same antigens is determined based on comparison with the negative and positive control.
The approach provides differential diagnosis by using spatially separated arbovirus antigens on one solid phase and then detecting and comparing corresponding signals. The documentation supports distinguishing patterns using direct comparison of Dengue virus NS1 band reactivity versus Zika virus NS1 band reactivity and identifying Chikungunya virus E1 band presence. It additionally emphasizes that IgG versus IgM reactivity can be used to interpret timing and that cross-reactivity effects are considered, including examples involving vaccination or other flavivirus exposure.
Claims Coverage
Independent claim clm-00001 defines an immunological differential diagnosis method with multiple spatially separated arbovirus antigens and signal comparison to negative and positive controls, determining IgG and/or IgM presence/absence.
Differential diagnosis with a spatially separated arbovirus antigen panel on a solid phase
A method for immunological differential diagnosis in which a sample is brought into contact with a plurality of antigens, separated from each other and applied to a solid phase, using at least Dengue virus serotype 2 NS1 and envelope protein antigens, Chikungunya virus envelope protein antigens, and Zika virus NS1 and envelope protein antigens, each as immunologically reactive parts having at least 40 amino acids.
Washing non-specific binding on the solid phase
The solid phase is washed to separate non-specific bindings.
Detection via anti-human IgG/IgM coupled to a signal-generating agent and immune-complex signal detection
Anti-human IgG and/or anti-human IgM antibodies coupled to a signal-generating agent are added to the washed solid phase, and any immune complex formed is detected by a signal from the signal-generating agent.
Signal comparison to negative and positive controls to determine IgG/IgM presence and/or absence
Any signal generated is compared to a negative and positive control, and the presence and/or absence of IgG and/or IgM antibodies against the same antigens is determined by comparing the signal with the negative and positive control.
The claims coverage centers on a differential diagnosis workflow that combines a spatially separated arbovirus antigen set on a solid phase, washing to remove non-specific binding, detection using anti-human IgG/IgM antibodies coupled to a signal-generating agent, and determination of antibody presence/absence by comparison to negative and positive controls.
Stated Advantages
Enables immunological differential diagnosis by determining the presence and/or absence of IgG and/or IgM antibodies against the same spatially separated arbovirus antigens using signals compared to negative and positive controls.
Allows IgG and/or IgM antibody detection directed to the same antigens used on the solid phase, supporting differentiation patterns across arbovirus antigens.
Documented Applications
Immunological differential diagnostic testing for arboviruses, including Dengue, Zika, and Chikungunya, with optional Yellow Fever NS1-derived antigen inclusion.
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