Viral detection method using viral encoded enzymes and chemiluminescent substrates

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.

1 . A method of detecting influenza virus in a subject suspected of being infected with influenza, said method comprising: 
 (1) obtaining a biological fluid sample from the subject;    (2) contacting at least a portion of the biological fluid sample with a substrate containing a chemiluminescent derivative having a formula                          wherein each R is independently selected from the group consisting of alkyl radicals have 1 to 6 carbon atoms, such that when the chemiluminescent derivative is in contact with influenza virus contained in the fluid sample, it will yield a chemiluminescence product which will generate light; and    (3) detecting whether light has been generated.    
   
   
       2 . The method as described in  claim 1 , wherein R is a methyl radical.  
   
   
       3 . The method as defined in  claim 2 , wherein a high or ultra-high speed photographic film is used to detect whether light has been generated.  
   
   
       4 . The method as defined in  claim 1 , wherein a high or ultra-high speed photographic film is used to detect whether light has been generated.  
   
   
       5 . The method as defined in  claim 1 , further comprising mixing the fluid sample with a diluent prior to contacting the chemiluminescent derivative.  
   
   
       6 . The method as defined in  claim 2 , further comprising mixing the fluid sample with a diluent prior to contacting the chemiluminescent derivative.  
   
   
       7 . The method as defined in  claim 1 , wherein the biological fluid sample is obtained from an oral cavity, a pharyngeal cavity, or a nasopharyngeal cavity.  
   
   
       8 . The method as defined in  claim 6 , wherein the biological fluid sample is obtained from an oral cavity, a pharyngeal cavity, or a nasopharyngeal cavity.