Mycoplasma Control

Intended use

The quality control of analytical methods is important in the maintenance of good clinical laboratory practices. Internal quality control in a microbiological laboratory requires the use of known strains. These microorganisms are frequently lyophilized in order to prolong shelf-life. The detection of urogenital mycoplasmas in clinical specimens is a routine laboratory test. The main mycoplasma species isolated from the urogenital tract are Ureaplasma urealyticum (U.u) and Mycoplasma hominis (M.h). These bacteria are present as commensal organisms, but can be pathogenic in nature.

Principle

Mycoplasmas are fragile germs lacking a cell wall and are demanding in their requirement for growth factors. The lyophilized U.u and M.h strains are produced from an enriched mycoplasma broth that contains excipients to protect the microorganisms during the lyophilization process.

Simple protocol

Allow the reagents to reach room temperature (18 to 25 °C) before use. Reconstitute the contents of the U.u or M.h vial with 1 mL of sterile distilled water (bacterial suspension). MYCOPLASMA CONTROL is validated for different commercialized methods. Please refer to the technical insert to follow the correct protocol and make the interpretation by comparison with the expected results. Please find below an example of methodology for our MYCOFAST^® EvolutioN 3 kit.

Example: MYCOFAST^® EvolutioN 3

Regenerate an UMMlyo (+ 3 mL UMMt) medium with an UMMt (3 mL) medium. Transfer 300 μL of the bacterial suspension into this medium (dilution at 1/10). Inoculate the MYCOFAST^® EvolutioN 3 tray as follows:

• 100 μL of inoculated UMM medium in each of the 20 wells
• 50 μL of S.Mh in wells 6 and 7
• 2 drops of paraffin oil in each of the 20 wells

Incubate the tray at 37 °C +/- 1 °C for 24 hours and interpret by comparison with the expected results:

Reagents and material

• MYCOPLASMA U.u:
  • 6 vials containing lyophilized U. urealyticum strain.
  • Before lyophilization each vial contains 0.5 mL of culture containing mycoplasma broth base, foal serum, mannitol and antibiotics. The U.u and M.h strains have been isolated from clinical specimens and characterized according to standard methods.
• MYCOPLASMA M.h:
  • 6 vials containing lyophilized M. hominis strain.
  • Before lyophilization each vial contains 0.5 mL of culture containing mycoplasma broth base, foal serum, mannitol and antibiotics. The U.u and M.h strains have been isolated from clinical specimens and characterized according to standard methods.

Stability and storage

• The strains stored at 2 to 8 °C in their original packaging, are stable until the expiry date shown on the kit.
• The vial of U.u. and M.h. strains should be reconstituted before use. The regenerated strains should be used immediately or stored for 24 hours at 2-8 °C maximum. Do not freeze.

Performance

• Identification / Purity
  • The U.u and M.h were identified on A7 agar plates and in U9 medium for U.u and Hayflick medium for M.h. There is no cross-contamination between U.u and M.h strains. Moreover there is no bacterial or mycological contamination on Columbia or Sabouraud agar plates.
• Enumeration
  • The U.u and M.h have been enumerated in a liquid medium by using the reference method (U9 medium for U.u and Shepard medium for M.h). The actual results obtained in the different trays agree with the expected results.
• Antibacterial susceptibility testing
  • The antibacterial susceptibility testing results obtained with the U.u and M.h in the different trays agree with the minimal inhibitory concentrations (MIC) in liquid media.
• Repeatability and Reproducibility
  • Studies carried out with two different U.u and M.h batches, for the control of MYCOFAST^® EvolutioN 3, demonstrated 100% repeatability and 100 % reproducibility within and between the batches. Studies carried out to test 10 different batches of MYCOFAST^® EvolutioN 2 tray, MYCOFAST^® EvolutioN 3 tray, UMMlyo (2 mL) and UMMlyo (3 mL) media, demonstrated that there is homogeneity in the results with U.u and M.h strains.

Material required but not provided

• Waste container for contaminated waste
• Sterile pipettes
• Sterile distilled water

Manufacturer: ELITech Microbio.
Product(s) intended for healthcare professionals.
Read the instructions on the label and/or instructions for use of the product(s).