Rapid Mycoplasma Test

€155.00*

Content: 1 unit
Services
Product number: 900159
Product nr Options Price
900159 Rapid Mycoplasma Test €155.00
900164 Premium Mycoplasma Test €360.00
Product information "Rapid Mycoplasma Test"

Mycoplasmas are small prokaryotic organisms that are too small to be visible in microscopy and cannot be eliminated by common antibiotic reagents. Mycoplasma may affect cellular growth, proliferation, and morphology of cell lines which may lead to unreliable experimental results. Approximately 15 to 35% of laboratory cell cultures are contaminated with mycoplasma. Therefore, frequent control is recommended to guarantee a mycoplasma-free environment. 

Analysis method

We offer Mycoplasma testing as a rapid variant or as a premium option. In the rapid test, samples are tested immediately after their arrival whereas, in the premium version, the cells are cultivated for two weeks without any antibitiotics and are then tested for Mycoplasma. Mycoplasma testing is performed using a two-point detection system with both the PlasmoTest™ - Mycoplasma Detection Kit (Invivogen) and the Certus QC – mycoADVANCED detection kit (Certus).

Colorimetric reporter assay

This test is a cell-based colorimetric assay. In the presence of mycoplasma, a reporter cell line induces a signaling cascade triggering a change of color in the medium from red to blue. The assay is performed in 96-well multiwell plates. Signals are detected in a microplate spectrophotometer at 620-655 nm. All mycoplasma and acholeplasma species, but also other contaminants in cell culture such as bacteria, are detected. The results are shown in a bar chart below:



Isothermal amplification

Isothermal amplification is a fast and reliable test based on isothermal amplification of mycoplasma-specific DNA combined with real-time detection using a DNA-intercalating dye. The assay can detect six of the most common species that account for >95% of contaminations: M. orale, M. hyorhinis, M. arginini, M. fermentans, M. hominis and A. laidlawii. Due to sequence homology, other mycoplasma species will be detected as well (M. pneumoniae, M. gallisepticum and M. synoviae). To identify whether the sample is mycoplasma positive or negative, the melting temperature (Tm) is studied. The results for the different melting temperatures are shown below: 



Well

Color

Sample Name

Tm (°C)

Results

1

Purple

Sample

94,65

Negative

2

 Pink 

Positive control

87,37

Positive

3

 Turquoise

Negative control

94,16

Negative

 

Glossary

Assessment of results of the Isothermal Amplification

All melting temperatures and their respective test results are explained below:

Sample

Melting temperature (Tm) in °C

Negative control (NC)/ internal control (IC)

94.8 +/- 0.2

Positive control (PC)

87.9 +/- 0.2 

Mycoplasma positive

- Amplification of mycoplasma DNA

87.0 - 89.2

Mycoplasma positive

- contaminated with a low amount of mycoplasma; Amplification of mycoplasma DNA and the internal control 

87.0 - 89.2 and 

95.0 +/- 0.2

Mycoplasma negative

- Amplification of the internal control

95.0 +/- 0.2

Contamination with other bacteria or unspecific 
 amplification

           89.5 - 94.0

 

Assessment of results of the Colorimetric Reporter Assay

If the test is positive, colony-forming units (CFU) in the range of 5*102 – 5*105 are present. A detailed list of the CFU/ml for different mycoplasma species is shown below:

 Mycoplasma species

Sensitivity CFU (ml)

A. laidlawii

2.5*103

M. arginini 

2.0*103

M. fermentans 

5.0*102

M. hyorhinis

7.0*102

M. orale 

5.0*105

 

Contamination-free cells
To identify mycoplasma contaminations we perform PCR-based and luminescence-based mycoplasma assays. We further determine any bacterial or fungal contamination through our standardized manufacturing processes.
Bulk order
Besides genomic DNA, RNA, cell pellets, and cell lysates in bulk, we can offer large quantities of assay-ready cells, plated cells in multiple formats, and frozen or growing cells. Contact us to receive a quote.
Authenticated cells
Each manufactured batch of cell lines* is authenticated via STR analysis. Contact us if you require a publication-ready STR report for your cells (*human, hamster, mouse, rat, and dog cells).
HLA alleles
HLA characterization is available from more than 200 cell lines. HLA class I -A, B, C, and Class II HLA-DPA1, -DPB1, -DQA1, DQB1, and DRB1 alleles were obtained by next-generation sequencing methodologies (NGS) for class I and class II alleles.