Mycoplasma pneumoniae immunological detection method and kit
[Problem] An object of the present invention is to provide a detection marker that can simply and rapidly detect Mycoplasma pneumoniae , which is a pathogen of mycoplasma pneumonia, at a high sensitivity, a specific antibody against the marker, and also an immunological detection method and a kit containing the antibody. [Solution] Infection with Mycoplasma pneumoniae can be rapidly and specifically diagnosed by producing an antibody specifically reactive to P30 protein of Mycoplasma pneumoniae and performing an immunological assay using the P30 protein as a detection marker. The present invention enables easy and rapid detection of Mycoplasma pneumoniae and diagnosis of infection with the bacteria at a hospital or the like without need of specialized instruments or skilled techniques.
1. An immunological method of detecting Mycoplasma pneumoniae , comprising subjecting a biological sample to a sandwich immunoassay using first and second antibodies against P30 protein of Mycoplasma pneumoniae to detect whether Mycoplasma pneumoniae is present in the biological sample or not, wherein at least one of the first and second antibodies is a monoclonal antibody recognizing an epitope of P30 protein present in a region of the amino acid sequence of SEQ ID NO: 2.
2. The method according to claim 1 , wherein the sandwich immunoassay is an ELISA or immunochromatographic assay.
3. The method according to claim 1 , wherein said monoclonal antibody recognizes a region having an amino acid sequence containing proline.
4. The method according to claim 1 , wherein one of the first and second antibodies is immobilized in a carrier.
5. An immunochromatographic assay for detecting Mycoplasma pneumoniae , comprising:
providing a membrane carrier having a capturing zone that is formed by previously immobilizing a first antibody against P30 protein of Mycoplasma pneumoniae at a predetermined position;
chromatographically developing a liquid mixture in the membrane carrier toward the capturing zone, said liquid mixture containing a second antibody against the P30 protein and a predetermined amount of a test sample,
whereby a complex of an antigen contained in the test sample and the second antibody is captured by the capturing zone,
wherein at least one of the first and second antibodies is a monoclonal antibody recognizing an epitope of P30 protein present in a region of the amino acid sequence of SEQ ID NO: 2, and
the test sample is a biological sample.
6. The immunochromatographic assay according to claim 5 , wherein said monoclonal antibody recognizes a region having an amino acid sequence containing proline.
7. The immunochromatographic assay according to claim 5 , herein the second antibody is labeled with a colloidal metal or a latex.
8. The immunochromatographic assay according to claim 7 , wherein the membrane carrier is a nitrocellulose membrane.
9. The method according to claim 1 , wherein the biological sample is at least one selected from the group consisting of nasal cavity aspirate, nasal cavity swab, throat swab, sputum, saliva, and bronchial washings.
10. The method according to claim 5 , wherein the biological sample is at least one selected from the group consisting of nasal cavity aspirate, nasal cavity swab, throat swab, sputum, saliva, and bronchial washings.