Human Mycoplasma IgG ELISA Kit

Sample

Human sera

Species Reactivity

Human

Detection Method

sELISA

Intended Use

The Mycoplasma IgG ELISA test system provides a means for the qualitative detection of IgG antibodies to Mycoplasma pneumoniae in human sera. The test may aid in the determination of the patient's serological status, or may aid in the diagnosis of disease associated with Mycoplasma pneumoniae. Potential cross-reactivity with M. genitalium has not been assessed, nor were studies performed on very young and/or elderly patients.

Contents of Kit

Each kit contains the following components in sufficient quantities to perform the number of tests indicated on packaging label. Note: All reactive reagents contain sodium azide as a preservative at a concentration of 0.1% (w/v).
1. Plate. 96 wells configured in twelve 1x8-well strips coated with partially purified inactivated M. pneumoniae (strain FN) 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 (γ 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 phosphatebuffered-saline, (pH 7.2 ± 0.2). Ready to use. Shake Well Before Use. NOTE: Sample Diluent will change color in the presence of serum.
7. TMB: One 15 mL amber bottle (amber cap) containing S.S'.S.S'-tetramethylbenzidine (TMB). Ready to use. Contains DMSO < 15% (w).
8. Stop Solution: One 15 mL bottle (red cap) containing 1M H₂SO₄, 0.7M HCI. Ready to use.
9. Wash Buffer concentrate (10X): dilute 1 part concentrate + 9 parts deionized or distilled water. One 100 mL bottle (clear cap) containing a 10X concentrated phosphate-buffered-saline and Tween-20 solution (blue solution). Note: 1X solution will have a pH of 7.2 ± 0.2.
Note: Kit also contains: Component list containing lot specific information is inside the kit box. Package insert providing instructions for use.

Storage

1. Store the unopened kit between 2°C and 8°C.
2. Microtiterwells: Store between 2°C and 8°C. Extra strips should be immediately resealed with desiccant and returned to proper storage. Strips are stable for 60 days after the envelope has been opened and properly resealed and the indicator strip on the desiccant pouch remains blue.
3. Conjugate: Store between 2°C and 8°C. DO NOT FREEZE.
4. Calibrator, Positive Control and Negative Control: Store between 2°C and 8°C.
5. TMB: Store between 2°C and 8°C.
6. Wash Buffer concentrate (10X): Store between 2°C and 25°C. Diluted wash buffer (1X) is stable at room temperature (20°C to 25° C) for up to 7 days or for 30 days between 2°C and 8°C.
7. Sample Diluent: Store between 2°C and 8°C.
8. Stop Solution: Store between 2°C and 25°C.

Performance Characteristics

Comparative Study:
A comparative study was performed to demonstrate the equivalence of the CD Mycoplasma IgG ELISA test system to a Crowntitre IgG IFA test system.
The performance of the CD Mycoplasma IgG ELISA test system was evaluated in a two site clinical investigation. There were a total of 194 specimens tested; 109 at site one, and 85 at site two. Most of the specimens (192/194) were obtained from a reference laboratory in Northeastern United States. These specimens were sent to the lab for routine Mycoplasma serological analysis. The remaining two specimens were repository specimens which had been previously tested for Mycoplasma IgG antibody, and were found to be positive. All specimens were frozen and maintained according to the guidelines indicated under the Specimen Collection section of this insert.
Specimens were tested on the ELISA test system at the clinical sites, and were then tested in-house by IFA. Table 1 below shows the results of this comparative study. These results represent those from single patient samples and not from multiple draws from the same patient.
Table 1: Calculation of Relative Sensitivity, Specificity, and Agreement

Relative Sensitivity = 69/73 = 94.5% (95% Confidence Interval* = 89.3 to 99.7%)
Relative Specificity = 84/96 = 87.5% (95% Confidence Interval* = 80.9 to 94.1%)
Relative Agreement = 153/169 = 90.5% (95% Confidence Interval* = 86.1 to 94.9%)
* 95% confidence intervals calculated using the exact method.
In addition to the two-site clinical study described above, the Mycoplasma IgG ELISA was used to evaluate 35 pairs of acute and convalescent specimens which were previously characterized by complement fixation (CF). Of the 35 pairs, 29 pairs demonstrated a four-fold or greater increase in the CF endpoint titer. Of the 29 pairs, 16 pairs were ELISA negative at the acute stage, and positive at the convalescent stage; eight pairs were positive at both the acute and convalescent stage; and five pairs were negative at both the acute and convalescent stage.
NOTE: Be advised that relative refers to the comparison of this assay's results to that of a similar assay. There was not an attempt to correlate the assay's results with disease presence or absence. No judgment can be made on the comparison assay's accuracy to predict disease.