Immunological detection method and kit for mycoplasma pneumoniae

Abstract

The present invention aims at providing a specific antibody that can simply and rapidly detect Mycoplasma pneumoniae which is a causative bacterium of mycoplasma pneumonia, with high sensitivity, and also an immunological detection method and a kit containing the same antibody. The present invention makes it possible to diagnose infection with Mycoplasma pneumoniae more rapidly and specifically than the conventional method, by producing an antibody recognizing a specific epitope of P30 protein of Mycoplasma pneumoniae and performing an immunological detection using the antibody. Also, the present invention enables easy and rapid detection of Mycoplasma pneumoniae and diagnosis of infection with the same at a hospital or the like without need of specialized instruments or skilled techniques.

1 . A method of detecting  Mycoplasma pneumoniae , comprising an immunoassay using an antibody against P30 protein of  Mycoplasma pneumoniae , the antibody being an antibody against an epitope of P30 protein located in any one of amino acid sequences of SEQ ID NOS: 3 to 5. 
     
     
         2 . The detecting method according to  claim 1 , wherein the antibody is a monoclonal antibody. 
     
     
         3 . A method of detecting  Mycoplasma pneumoniae , comprising a sandwich immunoassay using first and second antibodies against P30 protein of  Mycoplasma pneumoniae , wherein at least one of the first and second antibodies is an antibody against an epitope of P30 protein located in any one of amino acid sequences of SEQ ID NOS: 3 to 5. 
     
     
         4 . (canceled) 
     
     
         5 . The detecting method according to  claim 3 , wherein at least one of the first and second antibodies is a monoclonal antibody. 
     
     
         6 . The detecting method according to  claim 5 , wherein one of the first and second antibodies is immobilized in a carrier. 
     
     
         7 . (canceled) 
     
     
         8 . (canceled) 
     
     
         9 . (canceled) 
     
     
         10 . (canceled) 
     
     
         11 . An immunoassay kit for  Mycoplasma pneumoniae , comprising at least an antibody against P30 protein of  Mycoplasma pneumoniae , wherein the antibody is an antibody against an epitope of P30 protein located in any one of amino acid sequences of SEQ ID NOS: 3 to 5. 
     
     
         12 . A sandwich immunoassay kit for  Mycoplasma pneumoniae , comprising at least first and second antibodies against P30 protein of  Mycoplasma pneumoniae , wherein at least one of the first and second antibodies is an antibody against an epitope of P30 protein located in any one of amino acid sequences of SEQ ID NOS: 3 to 5. 
     
     
         13 . The sandwich immunoassay kit according to  claim 12 , wherein the sandwich immunoassay is an ELISA or immunochromatographic assay. 
     
     
         14 . A  Mycoplasma pneumoniae -detecting immunochromatographic test strip, comprising at least first and second antibodies against P30 protein of  Mycoplasma pneumoniae  and a membrane carrier,
 wherein the first antibody is previously immobilized at a predetermined position of the membrane carrier so as to form a capturing zone; and   the second antibody is labeled with an appropriate labeling substance and is provided at a position separated from the capturing zone so as to be chromatographically developed in the membrane carrier,   wherein at least one of the first and second antibodies is an antibody against an epitope of P30 protein located in any one of amino acid sequences of SEQ ID NOS: 3 to 5.   
     
     
         15 . The immunochromatographic test strip according to  claim 14 , wherein at least one of the first and second antibodies is a monoclonal antibody. 
     
     
         16 . The immunochromatographic test strip according to  claim 15 , wherein the second antibody is labeled with any one of a colloidal metal, a latex, and a fluorescent substance. 
     
     
         17 . The immunochromatographic test strip according to  claim 16 , wherein the membrane carrier is a nitrocellulose membrane. 
     
     
         18 . (canceled) 
     
     
         19 . (canceled) 
     
     
         20 . The detecting method according to  claim 2 , which exhibits a sensitivity sufficient for detecting  Mycoplasma pneumoniae  strain M129 in a culture having a concentration of 1×10 6  CFU/ml of the M129 strain. 
     
     
         21 . The detecting method according to  claim 5 , wherein the first and second antibodies, which may be the same or different from each other, are each a monoclonal antibody against an epitope of P30 protein located in any one of amino acid sequences of SEQ ID NOS: 3 to 5, said detecting method exhibiting a sensitivity sufficient for detecting  Mycoplasma pneumoniae  strain M129 in a culture having a concentration of 1×10 6  CFU/ml of the M129 strain. 
     
     
         22 . The immunoassay kit according to  claim 11 , wherein the antibody is a monoclonal antibody, said immunoassay kit exhibiting a sensitivity sufficient for detecting  Mycoplasma pneumoniae  strain M129 in a culture having a concentration of 1×10 6  CFU/ml of the M129 strain. 
     
     
         23 . The sandwich immunoassay kit according to  claim 12 , wherein the first and second antibodies, which may be the same or different from each other, are each a monoclonal antibody against an epitope of P30 protein located in any one of amino acid sequences of SEQ ID NOS: 3 to 5, said sandwich immunoassay kit exhibiting a sensitivity sufficient for detecting  Mycoplasma pneumoniae  strain M129 in a culture having a concentration of 1×10 6  CFU/ml of the M129 strain. 
     
     
         24 . The immunochromatographic test strip according to  claim 15 , wherein the first and second antibodies, which may be the same or different from each other, are each a monoclonal antibody against an epitope of P30 protein located in any one of amino acid sequences of SEQ ID NOS: 3 to 5, said immunochromatographic test strip exhibiting a sensitivity sufficient for detecting  Mycoplasma pneumoniae  strain M129 in a culture having a concentration of 1×10 6  CFU/ml of the M129 strain. 
     
     
         25 . The detecting method according to  claim 21 , wherein the sandwich immunoassay is an ELISA or immunochromatographic assay.