Mycoplasma Contamination Of Cell Cultures

Drexler, Hans G.; Uphoff, Cord C.

doi:10.1002/0471250570.spi054

Cited by 29 publications

(60 citation statements)

References 14 publications

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“…The primary sources of contamination with M. arginini and M. orale are sera used for the culture and laboratory personnel, respectively (Garner et al, 2000). A transfer of mycoplasmas from other cultures by using the same media and laboratory equipment is also possible (Drexler and Uphoff, 2000).…”

Section: Discussionmentioning

confidence: 99%

Detection of mycoplasma contamination in cell cultures and bovine sera

Dvořáková¹,

Valíček²,

Reichelova³

2005

Vet. Med. - Czech

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Contamination of cell cultures and sera used for animal virus propagation with mycoplasmas represents a serious problem, especially in virology. Therefore specific control measures must be used. To achieve this we introduced PCR for the detection of mycoplasma species in cell cultures and compared its results with ELISA and microbiological culture. Seven mycoplasma species which are the most common contaminants of cell lines (Mycoplasma arginini, M. fermentans, M. hyorhinis, M. bovis, M. orale, M. hominis, and Acholeplasma laidlawii) were used to verify the method. Then we assessed five selected cell lines and three bovine sera by the PCR, ELISA and culture methods and compared the results. PCR was positive for all of the mycoplasma species tested. ELISA kit used (Mycoplasma detection kit, Roche, Germany) allowed detection of only four species of contaminating mycoplasmas (Acholeplasma laidlawii, Mycoplasma arginini, M. hyorhinis, and M. orale). All the methods detected contamination of the VERO and RK13 cell lines. The agents of contamination were determined by the species-specific ELISA kit as Mycoplasma arginini and M. orale, respectively. Other cell lines and sera tested were not contaminated with mycoplasma. The results confirmed that the PCR method used in the present study is a sensitive, fast and specific detection method of mycoplasma contaminations and is suitable for routine mycoplasma detection in cell cultures and bovine sera.

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Section: Discussionmentioning

confidence: 99%

Detection of mycoplasma contamination in cell cultures and bovine sera

Dvořáková¹,

Valíček²,

Reichelova³

2005

Vet. Med. - Czech

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“…1 There are principally three variations of PCR-based mycoplasma detection: (1) detection of any mycoplasma infection (genus-specific PCR); (2) detection of single mycoplasma species (species-specific PCR); and (3) identification of the mycoplasma species with species-specific PCR or genus-specific PCR followed by sequencing of the PCR product or restriction fragment length polymorphism analysis (reviewed in Ref. 11).…”

Section: Discussionmentioning

confidence: 99%

“…1,8,11 In particular, PCR is not limited by the ability of an organism to grow in culture; in certain areas, this molecular nucleic acid amplification may eventually replace biological amplification (ie growth in artificial media), a feature of paramount importance considering the fastidious nature of some mycoplasma species. 4 Thus, given the high percentage of mycoplasma-contaminated cell lines (Figure 1) and the need for a simple and practical test, PCR should prove to be the state of the art technique for mycoplasma detection in cell cultures.…”

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Detection of mycoplasma in leukemia–lymphoma cell lines using polymerase chain reaction

Uphoff

Drexler

2002

Leukemia

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The specific, sensitive and reliable detection of mycoplasma contamination in cell cultures is an important part of mycoplasma control. We have sought to develop and validate a method for mycoplasma detection which is sensitive and accurate, but also practical in the sense of time spent, costs, and applicability in the standard laboratory; finally, the method should be suitable for screening large numbers of test specimens. To that end, we adapted a previously developed polymerase chain reaction (PCR) method for daily routine application. This single-step PCR uses a mixture of primers annealing to gene sequences coding for evolutionarily conserved 16S rRNA of different mycoplasma species, including the ones most commonly found in cell cultures. An internal control was introduced to exclude any false-negative tests resulting from technical PCR problems. This mycoplasma detection by PCR has been validated prospectively on 201 consecutive leukemia-lymphoma cell lines received at the institute over a 3-year period and on 118 initially positive cell lines after anti-mycoplasma treatment with antibiotics. The sensitivity (detection of true positives) of this PCR detection assay was 96% and the specificity (detection of true negatives) was also 96%, with positive and negative predictive values (probability of correct result) of 86% and 99%, respectively. PCR defined the mycoplasma status with 96% accuracy (detection of true positives and true negatives). Besides the high sensitivity and specificity, further attractive features of the PCR approach are the ease and speed with which large numbers of specimens can be tested. PCR mycoplasma analysis provides a readily available, quick and reliable test system with which to manage the important issue of mycoplasma contamination of cell lines.

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“…2,[5][6][7][8][9][10][11] The antigens examined on MUTZ-8 cells are listed in Table 1. The following viruses were examined by DNA and reverse transcriptase-polymerase chain reaction: Epstein-Barr virus (EBV), hepatitis B and C viruses (HBV, HCV), human immunodeficiency virus (HIV), and human T cell leukemia viruses (HTLV)-I/II.…”

Section: Characterization Of Mutz-8 Cellsmentioning

confidence: 99%

New acute myeloid leukemia-derived cell line: MUTZ-8 with 5q−

MacLeod

Meyer

et al. 2002

Leukemia

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The advent of continuous human leukemia-lymphoma cell lines as a rich resource of abundant, accessible and manipulable living cells has contributed significantly to a better understanding of the pathophysiology of hematopoietic tumors. We describe the establishment of the new continuous leukemia cell line MUTZ-8 from a patient with acute myeloid leukemia (AML): MUTZ-8 was derived from the peripheral blood of a 63-year-old woman with AML M4 (25 years after onset of myelodysplastic syndromes, MDS). DNA fingerprinting confirmed authenticity and derivation of the cell line. The immunoprofiling as determined by flow cytometry showed that MUTZ-8 is positive mainly for myeloid but also some monocytic and megakaryocytic markers, whereas it is negative for T cell, B cell and erythroid markers. The cell line is constitutively cytokine-dependent, proliferation requiring externally added cytokines. The cytokine response profiles showed a two-to 10-fold growth stimulation of the cells by various cytokines, whereas other cytokines led to growth inhibition. Cytogenetic analysis confirmed the common clonal derivation of the cell line and the malignant clone predominating at the times of sampling. MUTZ-8 displays a deletion of the 5q31 AML/MDS region effected by a non-reciprocal translocation, t(5;11)(q21;q10). The scientific utility of MUTZ-8 lies (1) in its cluster of pathognomonic cytogenetic alterations including a 5q31 breakpoint and (2) in its absolute cytokine dependency and proliferative response to various cytokines.

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Mycoplasma Contamination Of Cell Cultures

Cited by 29 publications

References 14 publications

Detection of mycoplasma contamination in cell cultures and bovine sera

Detection of mycoplasma contamination in cell cultures and bovine sera

Detection of mycoplasma in leukemia–lymphoma cell lines using polymerase chain reaction

New acute myeloid leukemia-derived cell line: MUTZ-8 with 5q−

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