Rapid Detection and Differentiation of the Major Mycoplasma Contaminants in Cell Cultures Using Real-Time PCR with SYBR Green I and Melting Curve Analysis
A quantitative real‐time polymerase chain reaction (PCR) procedure followed by melting curve analysis, using the green fluorescence dye SYBR Green I, was developed for rapid detection and differentiation of mycoplasma contaminants in cell cultures. This method showed that the detection of the target...
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| Veröffentlicht in: | Microbiology and immunology 2005-01, Vol.49 (9), p.859-863 |
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| container_title | Microbiology and immunology |
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| creator | Harasawa, Ryô Mizusawa, Hiroshi Fujii, Masashi Yamamoto, Junko Mukai, Hiroyuki Uemori, Takashi Asada, Kiyozo Kato, Ikunoshin |
| description | A quantitative real‐time polymerase chain reaction (PCR) procedure followed by melting curve analysis, using the green fluorescence dye SYBR Green I, was developed for rapid detection and differentiation of mycoplasma contaminants in cell cultures. This method showed that the detection of the target sequence was linear over a range from 104 to 10 colony‐forming units (CFU) of the mycoplasma cells. Analysis of the melting temperature of the PCR products allowed differentiation of the major mycoplasma contaminants. These results demonstrate that the protocol described in the present study can decrease the time to obtain reproducible results by simultaneous detection and differentiation of the Mycoplasma species contaminating cell cultures. |
| doi_str_mv | 10.1111/j.1348-0421.2005.tb03675.x |
| format | Article |
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This method showed that the detection of the target sequence was linear over a range from 104 to 10 colony‐forming units (CFU) of the mycoplasma cells. Analysis of the melting temperature of the PCR products allowed differentiation of the major mycoplasma contaminants. 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These results demonstrate that the protocol described in the present study can decrease the time to obtain reproducible results by simultaneous detection and differentiation of the Mycoplasma species contaminating cell cultures.</description><subject>Cells, Cultured - microbiology</subject><subject>melting curve</subject><subject>mycoplasma</subject><subject>Mycoplasma - classification</subject><subject>Mycoplasma - genetics</subject><subject>Mycoplasma - isolation & purification</subject><subject>Organic Chemicals - analysis</subject><subject>Polymerase Chain Reaction - methods</subject><subject>real-time PCR</subject><subject>SYBR Green I</subject><subject>Transition Temperature</subject><issn>0385-5600</issn><issn>1348-0421</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2005</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqVkc-O0zAQxiMEYsvCKyCLA7cEO06chBNLlpZKLay6RYiTNU3HrEviFNth21fhaXH_aDkifJnx-DffyPNF0StGExbOm03CeFbGNEtZklKaJ35FuSjyZPcoGj08PY5GlJd5nAtKL6Jnzm0oTYu0zJ5GF0ywIs0zNop-L2Cr1-QaPTZe94aACTetFFo0XsOx1ivi75DMYdNbMt83_bYF1wGpe-Oh0waMd0QbUmPbknpo_WDRkS9Om-9kgdDGS90huakX5F77O3L77f2CTCyiIdPjvDm2_sDWg_2F5MpAu3faPY-eKGgdvjjHy2g5_rCsP8azz5NpfTWLm5xVecyVENAATTGFlLN1rtZcVUUlxEoBF4LRslAF8CZlWQPAKqEoD0tKOSrWrPhl9Poku7X9zwGdl512TfgJGOwHJ0UZFsjy8p8gK0SeVZwH8O0JbGzvnEUlt1Z3YPeSUXlwUG7kwSZ5sEkeHJRnB-UuNL88TxlWHa7_tp4tC8C7E3CvW9z_h7ScT-fHNEjEJwntPO4eJMD-kKLggf36aRKy8ex6fFvJG_4HWVu7SQ</recordid><startdate>20050101</startdate><enddate>20050101</enddate><creator>Harasawa, Ryô</creator><creator>Mizusawa, Hiroshi</creator><creator>Fujii, Masashi</creator><creator>Yamamoto, Junko</creator><creator>Mukai, Hiroyuki</creator><creator>Uemori, Takashi</creator><creator>Asada, Kiyozo</creator><creator>Kato, Ikunoshin</creator><general>Blackwell Publishing Ltd</general><scope>BSCLL</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7T7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20050101</creationdate><title>Rapid Detection and Differentiation of the Major Mycoplasma Contaminants in Cell Cultures Using Real-Time PCR with SYBR Green I and Melting Curve Analysis</title><author>Harasawa, Ryô ; 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| ispartof | Microbiology and immunology, 2005-01, Vol.49 (9), p.859-863 |
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| language | eng |
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| source | Wiley Free Content; MEDLINE; Freely Accessible Japanese Titles; Wiley Online Library Journals Frontfile Complete; Alma/SFX Local Collection |
| subjects | Cells, Cultured - microbiology melting curve mycoplasma Mycoplasma - classification Mycoplasma - genetics Mycoplasma - isolation & purification Organic Chemicals - analysis Polymerase Chain Reaction - methods real-time PCR SYBR Green I Transition Temperature |
| title | Rapid Detection and Differentiation of the Major Mycoplasma Contaminants in Cell Cultures Using Real-Time PCR with SYBR Green I and Melting Curve Analysis |
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