Myco-Visible Mycoplasma LAMP Detection Kit

Mycoplasmas   are commonly found in research laboratories and unaffected by many of the antibiotics that have been used to control bacterial contamination in cell cultures, especially those that affect cell wall formation, such as penicillin. The lack of a cell wall also means that some cells may be able to pass through 0.2 μm filters. There have been several studies to determine the prevalence of mycoplasma contamination in cell cultures. Some have estimated that it could be as high as 80%, while studies in the USA suggest a lower contamination rate of 10-15%. An average figure between 25% and 50% worldwide seems quite plausible. This is a very significant problem for the industry and could lead to serious economic losses. Most cases of cell culture contamination are caused by a group of about six to eight species with human, porcine, or bovine hosts, of which Mycoplasma orale and Mycoplasma hyorrhinis are the most common.

Myco-Visible Mycoplasma LAMP detection kit is highly sensitive, specific, and rapid assay for the detection of mycoplasma contaminations in cell cultures. Myco-Visible Mycoplasma LAMP detection kit achieves this by targeting and amplifying the 16S rRNA coding region of the mycoplasma genome at loci that are well conserved in all Mollicutes, including Acholeplasma laidlawii, Mycoplasma arginini, Mycoplasma fermentans, Mycoplasma hominis, Mycoplasma hyorhinis, Mycoplasma orale, Mycoplasma Pneumoniae and Mycoplasma Salivarium. The detection procedure can be completed within 60 mins and requires as little as 10 16S rRNA copies or 10 fg of Mycoplasma genomic DNA per reaction. No cross-reactivity with other bacterial, fungal and mammalian DNA by Primer-BLAST. The test assay is easy to perform with no thermal cycler required. Positive results can be visualized by color changes from pink to yellow.

Result Interpretation

• Test result shall be interpreted as positive if the reaction solution turns yellow or orange.
• Test result shall be interpreted as negative if the reaction solution remains as pink.

Performance

Figure 1: A 10-fold serial dilution ranging from 10³ to 10 copies/µL was performed using six strains of Mycoplasma spp. The results demonstrated that the kit can detect as few as 10 copies of 16S rRNA or 10 fg of Mycoplasma genomic DNA.

 

Figure 2: Specificity test of Myco-Visible Mycoplasma LAMP detection Kit. Purified gDNA at 1 x 105 copies/reaction from 6 bacteria species that are genetically related to mycoplasma were tested. All samples showed a negative result, indicating no cross-reactivity.

Application Notes

Myco-visible Mycoplasma LAMP Detection Kit is a rapid, sensitive and specific nucleic acid isothermal amplification test optimized for the detection of Mycoplasma spp in cell culture.

Usage Statement

Research Use Only (RUO)