Method and kit for detecting influenza virus, and method for diagnosing influenza virus infection
Inventors
TABATA, Kazuhito • Noji, Hiroyuki • Urano, Yasuteru • Kamiya, Mako
Assignees
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Publication Number
US-12371752-B2
Publication Date
2025-07-29
Expiration Date
Abstract
As a technique for detecting an influenza virus with an improved accuracy, there is provided a method for detecting an influenza virus in a biological sample by using a first probe, which is decomposed by an influenza virus-derived neuraminidase and a bacterium-derived neuraminidase to generate an optically detectable signal, and a second probe, which is decomposed by the bacterium-derived neuraminidase to generate an optically detectable signal and not decomposed by the influenza virus-derived neuraminidase.
Core Innovation
The disclosure describes an improved influenza detection and diagnosis approach that distinguishes influenza virus-derived neuraminidase activity from bacterium-derived neuraminidase to avoid false positives. A biological sample is mixed with a first probe and a second probe, and each probe is decomposed by neuraminidase to generate an optically detectable signal. The first probe is decomposed by both influenza virus-derived neuraminidase and bacterium-derived neuraminidase, producing a first optically detectable signal.
The second probe is decomposed by bacterium-derived neuraminidase to generate a second optically detectable signal and is not decomposed by influenza virus-derived neuraminidase. The signals generated from the first probe and the second probe are optically discriminatorily detected, and presence or absence of the influenza virus is determined using the ratio of the intensity of the first-probe signal to the intensity of the second-probe signal compared with a predetermined value.
In the diagnostic implementation described, an optical discriminator enables detection of influenza infection by calling the subject infected when the intensity ratio is equal to or more than the predetermined value, and calling the subject not infected when the ratio is less than the predetermined value. The disclosure further describes diagnostic implementations using a digital system with micro-spaces or microdroplets, including an array with hydrophobic division walls, for quantitative, high-sensitivity detection.
Claims Coverage
The document includes three independent claims: a method for detecting an influenza virus, a kit for detecting an influenza virus, and a method for diagnosing whether a subject is infected or not. Across these claims, three main inventive features are defined: using two neuraminidase-discriminating probes with optically detectable signals, distinguishing influenza versus bacterium-derived neuraminidase by differential decomposition of the probes, and determining infection status by optically discriminatory detection of probe signals using an intensity ratio threshold.
Two neuraminidase-discriminating probes with optically detectable signals
A method comprising mixing a biological sample with a first probe and a second probe to generate optically detectable signals from decomposition by influenza virus-derived neuraminidase and bacterium-derived neuraminidase for the first probe, and decomposition by bacterium-derived neuraminidase and not by influenza virus-derived neuraminidase for the second probe.
Optically discriminatorily detecting first and second probe signals
Detecting signals generated from the first probe and the second probe, wherein the signal generated from the first probe and the signal generated from the second probe can be optically discriminatorily detected.
Intensity ratio threshold for infection presence/absence
Determining presence of the influenza virus when the ratio of an intensity of the signal generated from the first probe to an intensity of the signal generated from the second probe is equal to or more than a predetermined value, and detecting absence when the ratio is less than the predetermined value.
Influenza detecting kit with a second probe defined by formula (1)
A kit comprising a first probe decomposed by influenza virus-derived neuraminidase and bacterium-derived neuraminidase to generate an optically detectable signal, and a second probe decomposed by bacterium-derived neuraminidase to generate an optically detectable signal and not decomposed by influenza virus-derived neuraminidase, wherein the second probe is a compound represented by formula (1) or a salt thereof with defined substituent and integer parameters.
All independent claims rely on differential decomposition of a two-probe system by influenza virus-derived versus bacterium-derived neuraminidase to generate optically detectable signals, followed by optically discriminatory detection and an infection call based on an intensity ratio threshold. The kit claim further limits the second probe to the specified compound represented by formula (1) or a salt thereof.
Stated Advantages
Avoids false positives by discriminating influenza virus-derived neuraminidase activity from bacterium-derived neuraminidase.
Documented Applications
Detecting an influenza virus in a biological sample.
Diagnosing whether a subject is infected or not with an influenza virus.
Diagnostic implementations using a digital system with micro-spaces or microdroplets, including an array with hydrophobic division walls, for quantitative, high-sensitivity detection.
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