Adapting a conventional pcr assay for Toxoplasma gondii detection to real-time quantitative pcr including a competitive internal control
We have developed a quantitative PCR assay (LightCycler®) using the pair of primers JW58 and JW59 for the detection of the 35- fold repeated B1 gene of Toxoplasma gondii. This real-time PCR, using fluorescence resonance energy transfert (FRET) hybridization probes, allows the quantification of T. go...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
EDP Sciences
2007-06-01
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| Series: | Parasite |
| Subjects: | |
| Online Access: | http://dx.doi.org/10.1051/parasite/2007142149 |
| _version_ | 1797429164872040448 |
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| author | Brenier-Pinchart M.P. Morand-Bui V. Fricker-Hidalgo H. Equy V. Marlu R. Pelloux H. |
| author_facet | Brenier-Pinchart M.P. Morand-Bui V. Fricker-Hidalgo H. Equy V. Marlu R. Pelloux H. |
| author_sort | Brenier-Pinchart M.P. |
| collection | DOAJ |
| description | We have developed a quantitative PCR assay (LightCycler®) using the pair of primers JW58 and JW59 for the detection of the 35- fold repeated B1 gene of Toxoplasma gondii. This real-time PCR, using fluorescence resonance energy transfert (FRET) hybridization probes, allows the quantification of T. gondii with several technical requirements not previously described: i) an internal amplification control (co-amplified in a single tube with the same primers), ii) Uracil-N-Glycosylase and iii) a standard curve corresponding to a serial dilution from a calibrated suspension of T. gondii ranging from 40 to 4.106 parasites in one ml of amniotic fluid (1 to 105 T. gondii/PCR). In artificial samples, one parasite could be detected if at least three reactions were performed. |
| first_indexed | 2024-03-09T09:09:16Z |
| format | Article |
| id | doaj.art-a0b850669fb040e18363d34879006397 |
| institution | Directory Open Access Journal |
| issn | 1252-607X 1776-1042 |
| language | English |
| last_indexed | 2024-03-09T09:09:16Z |
| publishDate | 2007-06-01 |
| publisher | EDP Sciences |
| record_format | Article |
| series | Parasite |
| spelling | doaj.art-a0b850669fb040e18363d348790063972023-12-02T09:29:25ZengEDP SciencesParasite1252-607X1776-10422007-06-0114214915410.1051/parasite/2007142149parasite2007142p149Adapting a conventional pcr assay for Toxoplasma gondii detection to real-time quantitative pcr including a competitive internal controlBrenier-Pinchart M.P.Morand-Bui V.Fricker-Hidalgo H.Equy V.Marlu R.Pelloux H.We have developed a quantitative PCR assay (LightCycler®) using the pair of primers JW58 and JW59 for the detection of the 35- fold repeated B1 gene of Toxoplasma gondii. This real-time PCR, using fluorescence resonance energy transfert (FRET) hybridization probes, allows the quantification of T. gondii with several technical requirements not previously described: i) an internal amplification control (co-amplified in a single tube with the same primers), ii) Uracil-N-Glycosylase and iii) a standard curve corresponding to a serial dilution from a calibrated suspension of T. gondii ranging from 40 to 4.106 parasites in one ml of amniotic fluid (1 to 105 T. gondii/PCR). In artificial samples, one parasite could be detected if at least three reactions were performed.http://dx.doi.org/10.1051/parasite/2007142149Toxoplasma gondiiLightCycler®real-time PCRB1 geneinternal amplification control |
| spellingShingle | Brenier-Pinchart M.P. Morand-Bui V. Fricker-Hidalgo H. Equy V. Marlu R. Pelloux H. Adapting a conventional pcr assay for Toxoplasma gondii detection to real-time quantitative pcr including a competitive internal control Parasite Toxoplasma gondii LightCycler® real-time PCR B1 gene internal amplification control |
| title | Adapting a conventional pcr assay for Toxoplasma gondii detection to real-time quantitative pcr including a competitive internal control |
| title_full | Adapting a conventional pcr assay for Toxoplasma gondii detection to real-time quantitative pcr including a competitive internal control |
| title_fullStr | Adapting a conventional pcr assay for Toxoplasma gondii detection to real-time quantitative pcr including a competitive internal control |
| title_full_unstemmed | Adapting a conventional pcr assay for Toxoplasma gondii detection to real-time quantitative pcr including a competitive internal control |
| title_short | Adapting a conventional pcr assay for Toxoplasma gondii detection to real-time quantitative pcr including a competitive internal control |
| title_sort | adapting a conventional pcr assay for toxoplasma gondii detection to real time quantitative pcr including a competitive internal control |
| topic | Toxoplasma gondii LightCycler® real-time PCR B1 gene internal amplification control |
| url | http://dx.doi.org/10.1051/parasite/2007142149 |
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