A method for early nondestructive detection of mycoplasma contamination in cultured cells and its application

By using nucleic acid dyes and CFSE to label mycoplasma and combined with flow cytometry detection, the early non-destructive testing problem in the prior art was solved, and fast and accurate Mycoplasma pollution detection was achieved, reducing the detection time and risk of loss.

CN118834928BActive Publication Date: 2025-03-25SHENZHEN BAY LAB
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Patent Information

Application Number
CN202410920211.0
Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Filing Date
2024-07-10
Publication Date
2025-03-25
Estimated Expiration
2044-07-10

AI Technical Summary

Technical Problem

The prior art is difficult to detect the contamination of Mycoplasma in cultured cells in early and non-destructively, resulting in long-term detection, low accuracy, and may cause damage to cells.

Method used

Mycoplasma DNA is labeled using nucleic acid dyes such as Hoechst or DAPI, and intracellular protein amine groups are labeled in combination with CFSE dye, and then used flow cytometry to detect mycoplasma to achieve early non-destructive testing.

Benefits of technology

Fast, accurate and non-destructive Mycoplasma pollution detection is achieved, and results can be obtained in a very short time (1 to 2 hours), reducing the risk of economic and scientific research losses.

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Abstract

The present application discloses a method and application for early non-destructive detection of mycoplasma contamination in cultured cells. Mycoplasma DNA is labeled with nucleic acid dyes, intracellular protein amines are labeled with CFSE dyes, and then flow cytometry is used for detection. The developed method directly detects mycoplasma by labeling mycoplasma DNA and protein, especially when the mycoplasma density is low, the obtained results can be explained more clearly, and the early non-destructive detection of mycoplasma in cultured cells is truly realized. The present application provides a short-time, low-sampling, non-destructive, low detection concentration limit, and high-precision early detection method for mycoplasma contamination, which can detect and control mycoplasma contamination as early as possible, and minimize economic and scientific research losses.
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