In recent years, with the rapid development of biopharmaceuticals and the emergence of cell and gene therapies as well as mRNA vaccines during the pandemic, ensuring the safety and reliability of biological products has become a key focus and regulatory priority for governments and regulatory bodies worldwide. Mycoplasma is a common but often difficult-to-eliminate type of contamination. For bioprocesses involving cell culture, regulations require that 'there must be no mycoplasma contamination.
Points for Mycoplasma testing required by regulatory bodies
2.1 Cross-reactivity:
DNA from 14 strains of non-flexibacterial phyla and 6 engineering cells commonly used in biomedicine were selected, and none of the target channels peaked and the internal reference amplified normally.
|
Test Species |
|
|
Staphylococcus epidermidis |
Pseudohyphae |
|
Clostridium perfringens |
Salmonella enterica Enteritidis subsp. enterica |
|
Clostridium acetobutylicum |
Bacillus subtilis (bacillus subtilis) |
|
Acinetobacter baumannii (immobile bacteria) |
Bacillus cereus |
|
Lactobacillus acidophilus |
HEK293 |
|
Enterobacter aerogenes (taxonomic class of bacteria) |
Vero |
|
Micrococcus garciniae |
CHO |
|
streptococcus mutans |
E.coli |
|
Pseudomonas aeruginosa |
HEK293T |
|
Streptococcus pneumoniae |
Sf9 |
2.2 Some of the data are as follows:
- Detection Limit
Nucleic acid extraction and detection of 10CFU/mL Mycoplasma standard strains were performed according to the instructions of the standard strains and the extraction and detection kits.
The NCS and NTC samples were tested at the same time in each experiment, and the results of 24 replicate wells of 10CFU/mL samples of each strain met the requirement that ≥23 of the 24 tests were positive when the NCS and NTC results were satisfactory.
Table 1 Information on validated strains
|
Serial No. |
Mycoplasma name |
Concentration of bacterial liquid |
|
1 |
Mycoplasma arginini |
10 CFU/mL |
|
2 |
Mycoplasma orale |
10 CFU/mL |
|
3 |
Mycoplasma gallisepticum |
10 CFU/mL |
|
4 |
Mycoplasma pneumoniae |
10 CFU/mL |
|
5 |
Mycoplasma synoviae |
10 CFU/mL |
|
6 |
Mycoplasma fermentans |
10 CFU/mL |
|
7 |
Mycoplasma hyorhinis |
10 CFU/mL |
|
8 |
Acholeplasma laidlawii |
10 CFU/mL |
|
9 |
Spiroplasma citri |
10 CFU/mL |
|
10 |
Mycoplasma salivarium |
10 CFU/mL |
Table 2 Detection limit results
|
10 CFU/mL |
Experiment 1 |
Experiment 2 |
Experiment 3 |
|
Detection Rate |
8/8 |
8/8 |
8/8 |
|
Total |
24/24 |
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Product Information
|
Product |
Catalog Number |
Product Name |
product specification |
|
Sample pretreatment kit |
18461ES |
25T/100T |
|
|
18467ES |
MolPure® Mag48 Sample Preparation Kit FN |
3×16T/ 6×16T |
|
|
Nucleic acid extraction instrument |
80511ES |
48-Channel Automated Nucleic Acid Extractor |
48 Channel |
|
Mycoplasma Detection Kit |
40619ES |
25T/100T |