Viral detection method using viral encoded enzymes and chemiluminescent substrates

a technology of chemiluminescent substrates and enzymes, applied in enzymology, biochemistry apparatus and processes, instruments, etc., can solve the problem that the system is sufficiently unencumbered to be usable by a layperson

Inactive Publication Date: 2006-11-16
SHIMASAKI CRAIG D +2
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The system is sufficiently unencumbered as to be usable by a layperson in a nonmedical setting.

Method used

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  • Viral detection method using viral encoded enzymes and chemiluminescent substrates
  • Viral detection method using viral encoded enzymes and chemiluminescent substrates
  • Viral detection method using viral encoded enzymes and chemiluminescent substrates

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[0057] Throat swab specimens were collected using a foam swab from patients complaining of flu-like symptoms. The foam swab was then extracted into a buffered solution at pH 5.4 containing a detergent. The detergent extract of the patient specimen was then filtered through a 10μ filter. The filtrate was collected into amber vials containing the 1,2-dioxetane 4,7-dialkoxy-N-acetylneuraminic acid and then incubated at ambient temperature for 10 minutes. The reaction was terminated by adding a highly alkaline solution containing hydrophobic compounds and fluorescer molecules. This mixture was then transferred rapidly to a plastic manifold cell seated above a sensitive photographic film inside a light-tight chamber. Light from the released product undergoing decomposition strikes the photographic film to produce a white spot for image against a black background indicating the presence of either influenza A or influenza B virus in the patient specimen. Alternately, the emitted light from...

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Abstract

A chemiluminescent system for detecting the presence of influenza virus in a biological fluid sample is provided. An influenza diagnostic kit is provided which includes (1) a sampling device for obtaining the biological fluid from a subject, (2) a chemiluminescent substrate material which, in the presence of influenza virus in the biological sample, will generate a chemiluminescent product that will produce detectable light, and (3) a means for detecting any generated light. A liquid sample containing the biological fluid, and preferably a diluent, are contacted with the an absorbent material containing the chemiluminescent substrate material. The substrate responds to neuraminidase activity intrinsic to influenza A and influenza B virus particles, such that when the substrate is in contact with influenza virus, the substrate is cleaved to yield a chemiluminescent product that then decomposes to produce light which can then be detected. The chemiluminescent substrate materials include enzymatically triggerable 1,2-dioxetane derivatives of 4-alkoxy-N-acetylneuraminic acid and 4,7-dialkoxy-N-acetylneuraminic acid. The system is sufficiently simple that it can reliably be used by a layperson in a nonmedical setting. The biological fluid generally originates from the oral cavity, the pharyngeal cavity, or the nasopharyngeal cavity.

Description

RELATED APPLICATION [0001] This application is based on, and claims benefit of, U.S. Provisional Application Ser. No. 60 / 129,602, filed Apr. 16, 1999.FIELD OF THE INVENTION [0002] This invention relates to simple and direct systems and methods that permit the detection of influenza virus in a biological sample. The systems and methods of this invention are simple enough that they can reliably be used by non-medical personnel in a nonmedical setting. The invention provides an assay method that is simple, rapid and sensitive for use in detecting viruses, where an enzyme is encoded by the organism. It consists of a specimen collection step, an enzyme specific chemiluminescent substrate, and a high or ultra-high speed film unit for detection of that enzyme action on the specific chemiluminescent substrate. This method is especially adapted for detection and assay for influenza infections and viruses. BACKGROUND OF THE INVENTION [0003] Viral infections are a principal cause of illness du...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12Q1/70G01N33/483C12Q1/34G01N33/50G01N33/58
CPCC12Q1/34C12Q1/6813Y10S435/975C12Q1/70C12Q2563/103
Inventor SHIMASAKI, CRAIG D.ACHYUTHAN, KOMANDOOR ELAYAVALLIEDWARDS, BROOKS
Owner SHIMASAKI CRAIG D
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