Novel method for detecting mycoplasma infection in cell culture

Abstract

The invention belongs to the technical field of biological detection, and discloses a novel method for detecting mycoplasma infection in cell culture. A supernate of a cell culture solution is adoptdas a sample; extracted DNAs are adopted as a template to be added into a PCR system, and an amplified reaction is performed to obtain a PCR reaction product; nucleic acid removal water is adopted a template to be added into the PCR system, the amplified reaction is performed as negative control; after all the amplified reaction is ended, agarose gel electrophoresis is performed; an agarose gel electrophoresis detection map is observed; then, an ELISA reaction is performed; the sample concentration is determined. The experiment reliability and stability are ensured compared with independent setting of positive control. The PCR technology and the ELISA technology are combined, the rapid and sensitive advantages of the PCR technology are achieved, the method is better than the PCR method on the aspects of specificity identification and quantification, the experience result specificity is enhanced, the result is more credible objectively, and the defect that false positive and false negative results are likely to appear due to the PCR method is avoided.

Description

technical field [0001] The invention belongs to the technical field of biological detection, in particular to a new method for detecting mycoplasma infection in cell culture. Background technique [0002] Mycoplasma (Mycoplasma) is a class of prokaryotic microorganisms without cell walls between bacteria and viruses, and is the smallest microorganism known to be able to grow and reproduce in non-living media. The harm caused by mycoplasma infection is very extensive, and it can cause various diseases such as humans, animals, plants and insects, and has been widely paid attention to by people. However, due to the biological characteristics of mycoplasma such as small shape, diversity, slow growth and the existence of cross-antigens between mycoplasma species, some simple and intuitive methods for mycoplasma detection are limited. In the detection of mycoplasma by PCR method, due to the defects of PCR technology itself, there are also some problems, such as: prone to contamin...

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Problems solved by technology

In this way, the positive control must be set separately in the detection, which not only wastes reagents, but also makes the PCR process and electrophoresis detection process more cumbersome.

At the same time, the genomic DNA of mycoplasma must be prepared separately, which increases the complexity of the detection; the most important problem is that the detection system and the positive control system are in different PCR systems, even if the positive control is normal, there may be misjudgments

[0003] In summary, the existing problems in the prior art are: the existing mycoplasma detection method is prone to contamination, cross-reaction, false positive and false negative results, etc.; waste of reagents, PCR process and electrophoresis detection process will be more cumbersome, and errors may occur. Judgment

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