High Accuracy Mycoplasma Pneumoniae Rapid Test Elisa Sandwich Method CE

Product Specification

The Mycoplasma pneumoniae (M. pneumoniae) IgG ELISA, high accuracy, Elisa Sandwich method, for quantitative measurement

Product Name: The Mycoplasma pneumoniae (M. pneumoniae) IgG ELISA test

Intended Use:

Mycoplasma IgM Test System provides a means for the qualitative detection of IgM antibodies to Mycoplasma pneumoniae in human sera. When performed according to these instructions, the results of this test may aid in the diagnosis of M. pneumoniae infections in the adult population. This assay is for In Vitro diagnostic use only.

Summary:

Mycoplasma pneumoniae is a pathogen with spectrum of clinical presentations ranging from asymptomatic to pronounced pneumonia. Symptoms start from 6 to 32 days after exposure with headache, malaise, cough, sore throat and fever. The illness can last from a few days to a month or more. Detection by ELISA of M. pneumoniae IgM antibodies or demonstration of a significant increase of specific IgG antibodies is strong evidence for recent infection in the appropriate clinical setting. Specific IgM antibodies typically increase significantly 1 week after clinical onset and specific IgG levels rise in the second week. M. pneumoniae IgM can, however, persist for more than two years after infection, and therefore, detection of specific IgM does not accurately indicate the time of infection. Primary infection and reinfection may be distinguished by the presence of elevated specific IgA and of specific IgM in primary infections and by the presence of elevated specific IgA in the absence of specific IgM in reinfections. In general, the absence of specific IgM in serum collected 10-20 days after onset is strong evidence against primary pneumonia due to M. pneumoniae.

Regents Provided

Each kit contains the following components in sufficient quantities to perform the number of tests indicated on packaging label. Note: The following reactive reagents contain sodium azide as a preservative at a concentration of <0.1% (w/v): Controls, Sample Diluent, and Calibrator.

1.  Plate. 96 wells configured in twelve 1x8-well strips coated with inactivated preparation of M. pneumoniae (strain FH) antigen. The strips are packaged in a strip holder and sealed in an envelope with desiccant.
2.  Conjugate. Conjugated (horseradish peroxidase) goat anti-human IgG (Fc chain specific). Ready to use. One, 15 mL vial with a white cap.
3.  Positive Control (Human Serum). One, 0.35 mL vial with a red cap.
4.  Calibrator (Human Serum). One, 0.5 mL vial with a blue cap.
5.  Negative Control (Human Serum). One, 0.35 mL vial with a green cap.
6.  Sample Diluent. One 30 mL bottle (green cap) containing Tween-20, bovine serum albumin and phosphate-buffered-saline. Note: Sample Diluent will change color when combined with serum.

TEST PRINCIPLE

The Diagnostics Automation, Inc. Mycoplasma IgM ELISA test system is designed to detect IgM class antibodies to M. pneumoniae in human sera. Wells of plastic micro well strips are sensitized by passive absorption with M. pneumoniae antigen. The test procedure involves three incubation steps:

1.  Test sera are diluted with the Sample Diluent provided. The Sample Diluent contains anti-human IgG which precipitates and removes IgG and rheumatoid factor from the sample leaving IgM free to react with the immobilized antigen. During sample incubation, any antigen specific IgM antibody in the sample will bind to the immobilized antigen. The plate is washed to remove unbound antibody and other serum components.
2.  Peroxidase Conjugated goat anti-human IgM is added to the wells and the plate is incubated. The Conjugate will react with IgM antibody immobilized on the solid phase in step 1. The wells are washed to remove unbound Conjugate.
3.  The microwells containing immobilized peroxidase Conjugate are incubated with peroxidase Substrate Solution. Hydrolysis of the Substrate by peroxidase produces a color change. After a period of time the reaction is stopped and the color intensity of the solution is measured photometrically. The color intensity of the solution depends upon the antibody concentration in the original test sample.

TEST PROCEDURE

1.  Remove the individual components from storage and allow them to warm to room temperature (20-25°C).
2.  Determine the number of microwells needed. Allow six Control/Calibrator determinations (one Blank, one Negative Control, three Calibrators and one Positive Control) per run. A Reagent Blank should be run on each assay. Check software and reader requirements for the correct Controls/ Calibrator configurations. Return unused strips to the resealable pouch with desiccant, seal, and return to storage between 2°and 8°C.

1.  Prepare a 1:21 dilution (e.g.: 10µL of serum + 200µ L of Sample Diluent. of the Negative Control, Calibrator, Positive Control, and each patient serum.
2.  To individual wells, add 100mL of each diluted control, calibrator and sample. Ensure that the samples are properly mixed. Use a different pipette tip for each sample.
3.  Add 100µL of Sample Diluent to well A1 as a reagent blank. Check software and reader requirements for the correct reagent blank well configuration.
4.  Incubate the plate at room temperature (20-25°C) fo r 25 + 5 minutes.
5.  Wash the microwell strips 5X.