Product Name: Mycoplasma pneumoniae IgG - ELISA
# of Samples: 1 x 96 Assays
Intended Use: The M. pneumoniae IgG-ELISA is intended for the qualitative determination of IgG class antibodies against M. pneumoniae in human serum or plasma (citrate).
Introduction: The mycoplasms belong to the class Mollicutes comprising three distinct families and four genera, one of which is Mycoplasma with over 60 species. Mycoplasms are the smallest free living organisms known (300 to 500 nm in diameter) and unlike regular bacteria they lack a cell wall. Mycoplasms are extracellular parasites, especially on mucous membranes, which can cause infections in human, animals, plants, and cell cultures. Mycoplasma pneumoniae is primarily a respiratory pathogen (obligat) in human involving the nasopharynx, throat, trachea, bronchi, bronchioles, and alveoli. Other Mycoplasms, M. buccale, M. faucium, M. orale and M. salivarium are commensals in the oral cavity. Mycoplasma hominis and Ureaplasma urealyticum inhabit primarily the genital tract and may act as opportunistic invaders. M. pneumoniae is by far the most important pathogen of this group. Infection with M. pneumoniae occurs worldwide, its epidemiology has been studied primarily in the USA, Europe, and Japan. Infections are endemic in larger urban areas, and epidemic increases are observed at varying intervalls. M. pneumoniae has been estimated to cause 15-20% of all pneumoniae; the rate is highest in children and young adults. 74% of infections with M. pneumoniae are asymptomatic, reinfection may occur. Naturally acquired immunity to infection with M. pneumoniae appears to be of limited duration (2-3 years).
Infection may be identified by:
Hemadsorption, Tetrazolium reduction (specific for M. pneumoniae), Complement fixation
Detection of antibody production by ELISA Serology: complement fixation (CF), neutralization (N) and hemagglutination-inhibition (HAI); Detection of antibodies and the hexon antigen by ELISA.
Principles of the assay: The qualitative immunoenzymatic determination of IgG-class antibodies against M. pneumoniae is based on the ELISA (Enzyme-linked Immunosorbent Assay) technique.
Microtiter strip wells are precoated with M. pneumoniae antigens to bind corresponding antibodies of the specimen. After washing the wells to remove all unbound sample material horseradish peroxidase (HRP) labelled anti-human IgG conjugate is added. This conjugate binds to the captured M. pneumoniae-specific antibodies. The immune complex formed by the bound conjugate is visualized by adding Tetramethylbenzidine (TMB) substrate which gives a blue reaction product. The intensity of this product is proportional to the amount of M. pneumoniae-specific IgG antibodies in the specimen. Sulphuric acid is added to stop the reaction. This produces a yellow endpoint colour. Absorbance at 450 nm is read using an ELISA microwell plate reader
Storage and Stability: The reagents are stable up to the expiry date stated on the label when stored at 2...8 °C.
References: Clyde WA Jr:Clinical overviwe of typical M. pneumoniae infections.Clin.Infect.Dis 1993 Aug;17 Suppl 1:S32-6 MedlineE.Jacobs.1993.Serologicals Diagnosis of M. peumoniae Infections:Current Prodedures.Clinical Infectious Diseases.17(suppl 1) 79-82Foy,H.M.1993.Infections caused by M. pneumoniae and possible populations of patients.Clinical Infectious Diseases.17 (suppl 1) 37-46Handley J.G.,andL.D.Gray.1997.The incidence of M. pneumoniae Board Family Practice 11(6):425-429Vikerfors,T.,Brodin,G.,Grandien,M.,Hirshberg,L.,Krook,A.,andC.A.Petterssonspecific IgM antibodies for the diagnosis of M. pneumoniae infections Scand.J.Infect.Dis.20(6):601-610Wiegand,R.:M.pneumoniae.In:Brandis,H.,W.Köhler,H.J.Eggers,G.Pulverer(ed.):Med. Mikrobiologie 1994.Gustav Fischer Verlag Stuttgart,Jena,New York (1994) 767-769