สถาบันชีววัตถุ
กรมวิทยาศาสตร์การแพทย์
(National Control Laboratory for Biological Products)
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Annual Article 2006
Mycoplasmas are frequently contaminated in cultured cells. Without quality control of cells. In order to obtain cells with international standards for detection, direct detection can be performed by dyeing DNA with fluorescent colors in combination with culture in liquid and solid foods. The results showed that standard Mycoplasma pneumonia could be cultured well. Liquid and solid foods prepared in the laboratory in an oxygenated state. When laboratory cultures were first screened for Mycoplasmas, there was no evidence of contamination in the tested cells. However, the appropriate conditions for the cultivation of Mycoplasmas in the cell for positive control should be determined in conditions that mimic natural infection. Mycoplasma arginini and Acholeplasma laidlawii were ordered from ATCC to confirm that the cells in the laboratory were not contaminated with real Mycoplasmas or confirmed by PCR. The first research in the country to develop the detection of contamination of Mycoplasmas in the culture cell for a period of 2 years from 2005 to 2006.
Detection of Mycoplasmas Contamination in Culture Cells
Contamination of mycoplasma in cultured cells. One of the major problems in cell culture technology, therefore, continuous surveillance requires adequate testing. Currently, there are two main ways to confirm mycoplasma detection in cultured cells. By staining DNA with fluorescence. And DNA amplification by PCR was compared. Standardized methods of culture in solid and liquid foods The results showed that solid and liquid foods were prepared in the laboratory. Can promote the increase of the number of mycoplasma 3 species used. In this study, Mycoplasma pneumonia, Mycoplasma arginini, and Acholaplasma laidlawii were found. The three positive and two negative cells were negative by culture in solid foods for 21 days and by PCR for positive DNA from cultured surface cells. The sequential number of DNA sequences is shown at 191 bp with the mycoplasma DNA at 265-278 bp. In contrast, Hoechst 33258 indirect photocatalytic DNA analysis can not. Differentiate by always giving the same result. Culture in solid food To confirm that the PCR method is specific to detecting mycoplasma, the removal of mycoplasma from the cells was carried out. Used and found to negatively effect mycoplasma-negative DNA after disinfection. It has been concluded that DNA amplification by PCR is an appropriate method for routine screening and can be used in conjunction with Food culture for detection of mycoplasma contamination in cultured cells.
It is one of the big problems in the technology of cell culture. Consequently, continuous surveillance requires adequate testing methods. Currently, there are two main methods for confirming the detection of mycoplasma in cultured cells: indirectly by staining DNA with fluorescence and increasing Number of DNA by PCR method was compared.
Standard methods of culture in solid and liquid foods. The results showed that the solid and liquid foods used in the laboratory could increase the number of mycoplasma strains used in this study, namely Mycoplasma pneumonia, Mycoplasma arginini and Acholaplasma laidlawii. Surprisingly, the three positive surface species and the two suspensions were negative by culture in solid foods for 21 days and by PCR. For positive DNA from cultured surface cells, continuous expression of 191 bp internal control DNA was observed with the mycoplasma DNA strand at 265-278 bp. In contrast, DNA Hoechst 33258 cannot differentiate by giving the same result as the culture in solid foods. To confirm that the PCR method is specific to mycoplasma detection, the removal of mycoplasma from the cell was used and found to negatively effect mycoplasma-negative DNA after disinfection. It has been concluded that DNA amplification by PCR is an appropriate method for routine screening and can be used in combination with food culture for the detection of mycoplasma contamination in cultured cells.