|
There is No Trouble with Mycoplasma, and the Cells are Well Raised!
hits:46 Date:08/29/25
Mycoplasma is a common prokaryote that contaminates cells in laboratories. The main sources of contamination include cells and virus species, as well as animal raw materials such as serum, chicken embryos, primary cells and operators themselves. Data show that about 15%-35% of cells are contaminated by mycoplasma. Mycoplasma changes a series of characteristics such as the structure and function of host cells, resulting in inaccurate experimental results; Interfere with cell metabolism and growth, change nucleic acid synthesis, affect cell antigenicity, lead to chromosomal changes, interfere with virus replication and mimic the role of viruses. Therefore, the detection and removal of mycoplasma is very important for cell culture. It can be said that mycoplasma removal is indispensable in order to keep cells well.
The first challenge to remove mycoplasma contamination from cells is how to detect possible mycoplasma contamination in cells. Mycoplasma is a kind of prokaryotic microorganism with no cell wall and a diameter of about 0.1-0.3 μm. Conventional Gram staining is ineffective and difficult to observe under light microscope. Moreover, some mycoplasmas (such as Mycoplasma oralis) can establish latent infection with host cells without causing obvious cytopathic changes and are difficult to detect.
The classic methods for detecting mycoplasma are culture method and indicator cell method, but the two methods take too long, and the culture conditions are relatively harsh, so they are easily contaminated in the process, resulting in bias in the detection results. Based on the development of life science, there are some more efficient and accurate methods to replace traditional methods. As stated in < 2.6. 7 > of the European Pharmacopoeia, the nucleic acid amplification technique NAT can replace the culture method and indicator cell culture method after appropriate validation. The NAT method has high sensitivity and relatively short time consumption. The biochemical detection method based on the expression of mycoplasma-specific kinase takes less time, can complete the detection of mycoplasma within 20-30min, and its sensitivity is close to that of NAT method, which is more suitable for rapid detection and judgment of mycoplasma contamination.
| / |
Cultivation method |
Indicator cell method |
NAT (nucleic acid amplification technology) |
Biochemical method |
| Time |
28 days |
4-5 days |
2-5 hours |
20 minutes |
| Sensitivity |
10 cfu/mL |
100 cfu/mL |
10 cfu/mL |
50 cfu/mL |
| Specificity |
No single method achieves 100% specificity, and two methods are usually combined to improve specificity |
Table 1: Comparison of commonly used mycoplasma detection methods
Absin has launched two rapid detection products for mycoplasma detection, which are based on NAT and kinase biochemical detection technology respectively. The detection can be completed within half an hour, with a sensitivity of up to 10 cfu/mL, helping to rapidly detect mycoplasma contamination.
abs90062 one-step rapid mycoplasma test kit (isothermal amplification method) uses isothermal amplification technology and visual color changes to quickly detect 8 types of mycoplasma common in cell culture medium. The test can be completed within 30 minutes, no instruments are required.
Product Features:
1. Isothermal amplification technology, high sensitivity;
2. Simple operation, no PCR instrument and open-cover electrophoresis are required;
3. The experiment is fast and can be completed in 30 minutes;
4. There is no need for testing instruments, and the results are visualized;
abs90283 MYCO-LTM luciferase mycoplasma test kit can specifically detect kinases not expressed by eukaryotic cells produced by living mycoplasma, which can convert ADP to ATP. It can rapidly detect 8 common species and many other common live mycoplasmas within 30 minutes.
Product Features:
1. Bioluminescence detection, high sensitivity;
2. Simple operation, homogeneous reaction, two readings;
3. The experiment is fast and can be completed in 30 minutes;
4. High signal-to-noise ratio, comparable to the imported Lon * * brand;
Other mycoplasma testing products
| Item number |
Product name |
Specifications |
Principle characteristics |
| abs90062 |
One-step rapid mycoplasma test kit (isothermal amplification method) |
50T |
Isothermal amplification, 30min rapid detection, no PCR instrument required, visual results. |
| abs90283 |
MYCO-LTM Luciferase Mycoplasmic Test Kit |
10 & 25 & 50T |
Mycoplasma kinase luminescence method, the sensitivity is close to NAT method, and the rapid detection takes 20 minutes. |
| abs90199 |
Luminescence Mycoplasma Physical Test Kit |
20 & 200T |
Mycoplasma kinase luminescence method, 30 min to complete the experiment. |
| abs9252 |
Mycoplasma staining detection kit |
100T |
Hoechst staining detection, morphological observation |
| abs9588 |
Mycoplasma physical examination test kit (PCR method) |
50T |
NAT PCR detection, electrophoresis is required |
| abs9925 |
Mycoplasma test kit (qPCR method) |
50T |
NAT method, qPCR detection, Taqman probe, qPCR instrument real-time reading |
| abs90284 |
MYCO-T ™ Nested PCR Mycoplasma Physical Test Kit |
25 & 50T |
NAT method, nested PCR, two-step amplification, higher sensitivity, requires electrophoresis |
Mycoplasma pollution prevention and control needs to run through the entire process of "detection-removal-prevention"
Regular testing: It is recommended to conduct NAT or biochemical testing for precious cell lines once per passage;
Emergency treatment: immediately use abs9253 and other removal reagents after contamination is found, and cooperate with passage centrifugation to reduce mycoplasma load;
Environmental control: Use special disinfectants to regularly treat incubators, ultra-clean tables, and consumables such as serum require strict quality inspection.
Through professional testing products and standardized operation procedures, the mycoplasma protective barrier of cell culture can be effectively constructed to ensure the stability and reliability of scientific research experiments.
|
中文
