Mycoplasma contamination in human leukemia cell lines. I: Comparison of various detection methods
The sensitivity and reliability of seven assays for mycoplasma detection were tested on a panel of leukemia cell lines. The assays used were: microbiological cultivation on broth and agar, immunofluorescent visualization of mycoplasmal DNA using DAPI (both direct staining and after multiplication of...
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| Veröffentlicht in: | Journal of immunological methods 1992-04, Vol.149 (1), p.43-53 |
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| container_title | Journal of immunological methods |
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| creator | UPHOFF, C. C GIGNAC, S. M DREXLER, H. G |
| description | The sensitivity and reliability of seven assays for mycoplasma detection were tested on a panel of leukemia cell lines. The assays used were: microbiological cultivation on broth and agar, immunofluorescent visualization of mycoplasmal DNA using DAPI (both direct staining and after multiplication of the contaminants on an indicator cell line), a nucleic acid hybridization assay with a radioactive probe specific for mycoplasmal rRNA, and ELISA with mycoplasma-specific polyclonal antibodies, a biochemical method utilizing 6-MPDR, and a mycoplasma-specific monoclonal antibody in immunofluorescence staining. The broth-agar method, the two DAPI tests and the RNA hybridization assay produced the highest detection rates; a number of false-negative cases were recorded by the other tests. The detection rates, costs, requirement for specialized equipment and other characteristics were evaluated for each method. Since each technique also has disadvantages and certain limitations and since no method can be regarded as the 'gold standard', at least two procedures should be used in routine screening for mycoplasma in cell cultures. |
| doi_str_mv | 10.1016/S0022-1759(12)80047-0 |
| format | Article |
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The broth-agar method, the two DAPI tests and the RNA hybridization assay produced the highest detection rates; a number of false-negative cases were recorded by the other tests. The detection rates, costs, requirement for specialized equipment and other characteristics were evaluated for each method. 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C</creatorcontrib><creatorcontrib>GIGNAC, S. M</creatorcontrib><creatorcontrib>DREXLER, H. G</creatorcontrib><title>Mycoplasma contamination in human leukemia cell lines. I: Comparison of various detection methods</title><title>Journal of immunological methods</title><addtitle>J Immunol Methods</addtitle><description>The sensitivity and reliability of seven assays for mycoplasma detection were tested on a panel of leukemia cell lines. The assays used were: microbiological cultivation on broth and agar, immunofluorescent visualization of mycoplasmal DNA using DAPI (both direct staining and after multiplication of the contaminants on an indicator cell line), a nucleic acid hybridization assay with a radioactive probe specific for mycoplasmal rRNA, and ELISA with mycoplasma-specific polyclonal antibodies, a biochemical method utilizing 6-MPDR, and a mycoplasma-specific monoclonal antibody in immunofluorescence staining. The broth-agar method, the two DAPI tests and the RNA hybridization assay produced the highest detection rates; a number of false-negative cases were recorded by the other tests. The detection rates, costs, requirement for specialized equipment and other characteristics were evaluated for each method. Since each technique also has disadvantages and certain limitations and since no method can be regarded as the 'gold standard', at least two procedures should be used in routine screening for mycoplasma in cell cultures.</description><subject>Antibodies, Monoclonal</subject><subject>Biological and medical sciences</subject><subject>Cell Line</subject><subject>Cells, Cultured</subject><subject>DNA - analysis</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Fluorescent Antibody Technique</subject><subject>Fluorescent Dyes</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fundamental immunology</subject><subject>Humans</subject><subject>Immunobiology</subject><subject>Immunological reactions in vitro</subject><subject>Indoles</subject><subject>Leukemia - microbiology</subject><subject>Methods</subject><subject>Mycoplasma</subject><subject>Mycoplasma - genetics</subject><subject>Mycoplasma Infections - diagnosis</subject><subject>Nucleic Acid Hybridization</subject><subject>Purine Nucleosides</subject><subject>Reproducibility of Results</subject><subject>RNA - analysis</subject><subject>Sensitivity and Specificity</subject><issn>0022-1759</issn><issn>1872-7905</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1992</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE1LxDAQhoMo67r6ExZyENFD13w1ab3J4sfCigf1XGbTlI02aW1aYf-9WS1ePc3A-_DyzCA0p2RBCZXXL4QwllCV5peUXWWECJWQAzSlmWKJykl6iKZ_yDE6CeGdEEKJJBM0oVwJpfIpgqedbtoaggOsG9-Dsx5623hsPd4ODjyuzfBhnI25qWtcW2_CAq9u8LJxLXQ2RLap8FdcmyHg0vRG_xQ402-bMpyiowrqYM7GOUNv93evy8dk_fywWt6uE83SrE-qzHCe87SiWkAmKJW5hDxqghTASCUM1zQT-SajrExNlmpGuIi3RqAikZyhi9_etms-BxP6wtmwVwZvolihWC4kU-pfkEqWCs72jfMRHDbOlEXbWQfdrhifF_PzMYegoa468NqGPyxlTCrB-Td39337</recordid><startdate>19920427</startdate><enddate>19920427</enddate><creator>UPHOFF, C. 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Psychology</topic><topic>Fundamental immunology</topic><topic>Humans</topic><topic>Immunobiology</topic><topic>Immunological reactions in vitro</topic><topic>Indoles</topic><topic>Leukemia - microbiology</topic><topic>Methods</topic><topic>Mycoplasma</topic><topic>Mycoplasma - genetics</topic><topic>Mycoplasma Infections - diagnosis</topic><topic>Nucleic Acid Hybridization</topic><topic>Purine Nucleosides</topic><topic>Reproducibility of Results</topic><topic>RNA - analysis</topic><topic>Sensitivity and Specificity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>UPHOFF, C. C</creatorcontrib><creatorcontrib>GIGNAC, S. M</creatorcontrib><creatorcontrib>DREXLER, H. 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| ispartof | Journal of immunological methods, 1992-04, Vol.149 (1), p.43-53 |
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| language | eng |
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| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | Antibodies, Monoclonal Biological and medical sciences Cell Line Cells, Cultured DNA - analysis Enzyme-Linked Immunosorbent Assay Fluorescent Antibody Technique Fluorescent Dyes Fundamental and applied biological sciences. Psychology Fundamental immunology Humans Immunobiology Immunological reactions in vitro Indoles Leukemia - microbiology Methods Mycoplasma Mycoplasma - genetics Mycoplasma Infections - diagnosis Nucleic Acid Hybridization Purine Nucleosides Reproducibility of Results RNA - analysis Sensitivity and Specificity |
| title | Mycoplasma contamination in human leukemia cell lines. I: Comparison of various detection methods |
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