Prenatal diagnosis of congenital toxoplasmosis by duplex real-time PCR using fluorescence resonance energy transfer hybridization probes
The diagnosis of congenital toxoplasmosis frequently relies on PCR tests of amniotic fluid (AF). A duplex real‐time quantitative PCR test based on fluorescence resonance energy transfer was developed to quantify the parasite load and to decrease the risk of contamination. An internal control based o...
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| Veröffentlicht in: | Prenatal diagnosis 2001-02, Vol.21 (2), p.85-88 |
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| creator | Costa, Jean-Marc Ernault, Pauline Gautier, Evelyne Bretagne, Stéphane |
| description | The diagnosis of congenital toxoplasmosis frequently relies on PCR tests of amniotic fluid (AF). A duplex real‐time quantitative PCR test based on fluorescence resonance energy transfer was developed to quantify the parasite load and to decrease the risk of contamination. An internal control based on the detection of 10 pg mouse DNA added to the AF was included to check for PCR efficiency. The relationship between the parasite load and the occurrence of ultrasonographic abnormalities in 87 samples of AF was analyzed. Seven AF (8%) had a parasitic load >103; 14 (16%) had >102–≤103; 26 (30%) had >10–≤102; and 40 (46%) had ≤10 parasites/ml. Four of the six AF with cerebral ventriculomegaly had >103 parasites/ml. The other two had 130 and 24 parasites/ml, respectively. No parasitic loads of >103 parasites/ml and no ultrasonographic abnormalities were observed in the 11 AF with maternal toxoplasmosis in the third trimester. Therefore, there is a trend to associate high parasite count with ultrasonographic abnormality, but the main concern remains early maternal infection. The importance of quantification should be better evaluated with postnatal studies. The duplex LightCycler PCR test currently provides rapid and safe results. Copyright © 2001 John Wiley & Sons, Ltd. |
| doi_str_mv | 10.1002/1097-0223(200102)21:2<85::AID-PD18>3.0.CO;2-1 |
| format | Article |
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A duplex real‐time quantitative PCR test based on fluorescence resonance energy transfer was developed to quantify the parasite load and to decrease the risk of contamination. An internal control based on the detection of 10 pg mouse DNA added to the AF was included to check for PCR efficiency. The relationship between the parasite load and the occurrence of ultrasonographic abnormalities in 87 samples of AF was analyzed. Seven AF (8%) had a parasitic load >103; 14 (16%) had >102–≤103; 26 (30%) had >10–≤102; and 40 (46%) had ≤10 parasites/ml. Four of the six AF with cerebral ventriculomegaly had >103 parasites/ml. The other two had 130 and 24 parasites/ml, respectively. No parasitic loads of >103 parasites/ml and no ultrasonographic abnormalities were observed in the 11 AF with maternal toxoplasmosis in the third trimester. Therefore, there is a trend to associate high parasite count with ultrasonographic abnormality, but the main concern remains early maternal infection. The importance of quantification should be better evaluated with postnatal studies. The duplex LightCycler PCR test currently provides rapid and safe results. Copyright © 2001 John Wiley & Sons, Ltd.</description><identifier>ISSN: 0197-3851</identifier><identifier>EISSN: 1097-0223</identifier><identifier>DOI: 10.1002/1097-0223(200102)21:2<85::AID-PD18>3.0.CO;2-1</identifier><identifier>PMID: 11241531</identifier><identifier>CODEN: PRDIDM</identifier><language>eng</language><publisher>Chichester, UK: John Wiley & Sons, Ltd</publisher><subject>Amniocentesis ; Animals ; Antibodies, Protozoan - analysis ; Biological and medical sciences ; congenital toxoplasmosis ; Diseases of mother, fetus and pregnancy ; DNA, Protozoan - analysis ; Female ; Gynecology. Andrology. Obstetrics ; Human protozoal diseases ; Humans ; Infectious diseases ; Medical sciences ; Mice ; Nucleic Acid Hybridization ; Parasitic diseases ; PCR ; Polymerase Chain Reaction - methods ; Pregnancy ; Pregnancy. Fetus. Placenta ; prenatal diagnosis ; Prenatal Diagnosis - methods ; Protozoal diseases ; Spectrometry, Fluorescence ; Toxoplasma - genetics ; Toxoplasma - immunology ; Toxoplasmosis ; Toxoplasmosis, Congenital - diagnosis ; Ultrasonography, Prenatal</subject><ispartof>Prenatal diagnosis, 2001-02, Vol.21 (2), p.85-88</ispartof><rights>Copyright © 2001 John Wiley & Sons, Ltd.</rights><rights>2001 INIST-CNRS</rights><rights>Copyright 2001 John Wiley & Sons, Ltd.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c4148-4c4b7f3cde399e1c5543fdb9918e9c64175d50d0fa37cbf4d328aba62e58ba7e3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2F1097-0223%28200102%2921%3A2%3C85%3A%3AAID-PD18%3E3.0.CO%3B2-1$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2F1097-0223%28200102%2921%3A2%3C85%3A%3AAID-PD18%3E3.0.CO%3B2-1$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,27923,27924,45573,45574</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=891922$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11241531$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Costa, Jean-Marc</creatorcontrib><creatorcontrib>Ernault, Pauline</creatorcontrib><creatorcontrib>Gautier, Evelyne</creatorcontrib><creatorcontrib>Bretagne, Stéphane</creatorcontrib><title>Prenatal diagnosis of congenital toxoplasmosis by duplex real-time PCR using fluorescence resonance energy transfer hybridization probes</title><title>Prenatal diagnosis</title><addtitle>Prenat. Diagn</addtitle><description>The diagnosis of congenital toxoplasmosis frequently relies on PCR tests of amniotic fluid (AF). A duplex real‐time quantitative PCR test based on fluorescence resonance energy transfer was developed to quantify the parasite load and to decrease the risk of contamination. An internal control based on the detection of 10 pg mouse DNA added to the AF was included to check for PCR efficiency. The relationship between the parasite load and the occurrence of ultrasonographic abnormalities in 87 samples of AF was analyzed. Seven AF (8%) had a parasitic load >103; 14 (16%) had >102–≤103; 26 (30%) had >10–≤102; and 40 (46%) had ≤10 parasites/ml. Four of the six AF with cerebral ventriculomegaly had >103 parasites/ml. The other two had 130 and 24 parasites/ml, respectively. No parasitic loads of >103 parasites/ml and no ultrasonographic abnormalities were observed in the 11 AF with maternal toxoplasmosis in the third trimester. Therefore, there is a trend to associate high parasite count with ultrasonographic abnormality, but the main concern remains early maternal infection. The importance of quantification should be better evaluated with postnatal studies. The duplex LightCycler PCR test currently provides rapid and safe results. Copyright © 2001 John Wiley & Sons, Ltd.</description><subject>Amniocentesis</subject><subject>Animals</subject><subject>Antibodies, Protozoan - analysis</subject><subject>Biological and medical sciences</subject><subject>congenital toxoplasmosis</subject><subject>Diseases of mother, fetus and pregnancy</subject><subject>DNA, Protozoan - analysis</subject><subject>Female</subject><subject>Gynecology. Andrology. Obstetrics</subject><subject>Human protozoal diseases</subject><subject>Humans</subject><subject>Infectious diseases</subject><subject>Medical sciences</subject><subject>Mice</subject><subject>Nucleic Acid Hybridization</subject><subject>Parasitic diseases</subject><subject>PCR</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Pregnancy</subject><subject>Pregnancy. Fetus. Placenta</subject><subject>prenatal diagnosis</subject><subject>Prenatal Diagnosis - methods</subject><subject>Protozoal diseases</subject><subject>Spectrometry, Fluorescence</subject><subject>Toxoplasma - genetics</subject><subject>Toxoplasma - immunology</subject><subject>Toxoplasmosis</subject><subject>Toxoplasmosis, Congenital - diagnosis</subject><subject>Ultrasonography, Prenatal</subject><issn>0197-3851</issn><issn>1097-0223</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2001</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkcGO0zAQhiMEYkvhFZAlJASHFI8dN0lBSKsUdlcq2woW7dFynEkxmzrFTkTLE_DYONuqXDhwsj3-9M1o_igSQCdAKXsDNE9jyhh_xSgFyl4zmLF3mZjNzq_m8WoO2Xs-oZNi-ZbF8CAanfiH0YhCuPNMwFn0xPvvQZexPH0cnQGwBASHUfR75dCqTjWkMmptW288aWuiW7tGa4Z61-7abaP85v6v3JOq3za4Iw5VE3dmg2RVfCa9N3ZN6qZvHXqNVmMAfGvVcEOLbr0nnVPW1-jIt33pTGV-qc60lmxdW6J_Gj2qVePx2fEcR18_frgpLuPF8uKqOF_EOoEkixOdlGnNdYU8zxG0EAmvqzLPIcNcTxNIRSVoRWvFU13WScVZpko1ZSiyUqXIx9HLgze0_dGj7-TGhIGbRllsey_TaT5lkOUB_HQAtWu9d1jLrTMb5fYSqByikcOq5bBqeYhGMpBMZkLKEI0copFcUlksQxWC7_mxcV9usPprO2YRgBdHQHmtmjpsSxt_4rIc8tBrHF0fqJ-mwf3_z_SPke7fQRgfhMZ3uDsJlbuT05SnQt5eX8h58WXBLm-FvOF_ABoQxF0</recordid><startdate>200102</startdate><enddate>200102</enddate><creator>Costa, Jean-Marc</creator><creator>Ernault, Pauline</creator><creator>Gautier, Evelyne</creator><creator>Bretagne, Stéphane</creator><general>John Wiley & Sons, Ltd</general><general>Wiley</general><scope>BSCLL</scope><scope>IQODW</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>7X8</scope></search><sort><creationdate>200102</creationdate><title>Prenatal diagnosis of congenital toxoplasmosis by duplex real-time PCR using fluorescence resonance energy transfer hybridization probes</title><author>Costa, Jean-Marc ; Ernault, Pauline ; Gautier, Evelyne ; Bretagne, Stéphane</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4148-4c4b7f3cde399e1c5543fdb9918e9c64175d50d0fa37cbf4d328aba62e58ba7e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2001</creationdate><topic>Amniocentesis</topic><topic>Animals</topic><topic>Antibodies, Protozoan - analysis</topic><topic>Biological and medical sciences</topic><topic>congenital toxoplasmosis</topic><topic>Diseases of mother, fetus and pregnancy</topic><topic>DNA, Protozoan - analysis</topic><topic>Female</topic><topic>Gynecology. Andrology. Obstetrics</topic><topic>Human protozoal diseases</topic><topic>Humans</topic><topic>Infectious diseases</topic><topic>Medical sciences</topic><topic>Mice</topic><topic>Nucleic Acid Hybridization</topic><topic>Parasitic diseases</topic><topic>PCR</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Pregnancy</topic><topic>Pregnancy. Fetus. Placenta</topic><topic>prenatal diagnosis</topic><topic>Prenatal Diagnosis - methods</topic><topic>Protozoal diseases</topic><topic>Spectrometry, Fluorescence</topic><topic>Toxoplasma - genetics</topic><topic>Toxoplasma - immunology</topic><topic>Toxoplasmosis</topic><topic>Toxoplasmosis, Congenital - diagnosis</topic><topic>Ultrasonography, Prenatal</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Costa, Jean-Marc</creatorcontrib><creatorcontrib>Ernault, Pauline</creatorcontrib><creatorcontrib>Gautier, Evelyne</creatorcontrib><creatorcontrib>Bretagne, Stéphane</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Prenatal diagnosis</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Costa, Jean-Marc</au><au>Ernault, Pauline</au><au>Gautier, Evelyne</au><au>Bretagne, Stéphane</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Prenatal diagnosis of congenital toxoplasmosis by duplex real-time PCR using fluorescence resonance energy transfer hybridization probes</atitle><jtitle>Prenatal diagnosis</jtitle><addtitle>Prenat. Diagn</addtitle><date>2001-02</date><risdate>2001</risdate><volume>21</volume><issue>2</issue><spage>85</spage><epage>88</epage><pages>85-88</pages><issn>0197-3851</issn><eissn>1097-0223</eissn><coden>PRDIDM</coden><abstract>The diagnosis of congenital toxoplasmosis frequently relies on PCR tests of amniotic fluid (AF). A duplex real‐time quantitative PCR test based on fluorescence resonance energy transfer was developed to quantify the parasite load and to decrease the risk of contamination. An internal control based on the detection of 10 pg mouse DNA added to the AF was included to check for PCR efficiency. The relationship between the parasite load and the occurrence of ultrasonographic abnormalities in 87 samples of AF was analyzed. Seven AF (8%) had a parasitic load >103; 14 (16%) had >102–≤103; 26 (30%) had >10–≤102; and 40 (46%) had ≤10 parasites/ml. Four of the six AF with cerebral ventriculomegaly had >103 parasites/ml. The other two had 130 and 24 parasites/ml, respectively. No parasitic loads of >103 parasites/ml and no ultrasonographic abnormalities were observed in the 11 AF with maternal toxoplasmosis in the third trimester. Therefore, there is a trend to associate high parasite count with ultrasonographic abnormality, but the main concern remains early maternal infection. The importance of quantification should be better evaluated with postnatal studies. The duplex LightCycler PCR test currently provides rapid and safe results. Copyright © 2001 John Wiley & Sons, Ltd.</abstract><cop>Chichester, UK</cop><pub>John Wiley & Sons, Ltd</pub><pmid>11241531</pmid><doi>10.1002/1097-0223(200102)21:2<85::AID-PD18>3.0.CO;2-1</doi><tpages>4</tpages></addata></record> |
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| subjects | Amniocentesis Animals Antibodies, Protozoan - analysis Biological and medical sciences congenital toxoplasmosis Diseases of mother, fetus and pregnancy DNA, Protozoan - analysis Female Gynecology. Andrology. Obstetrics Human protozoal diseases Humans Infectious diseases Medical sciences Mice Nucleic Acid Hybridization Parasitic diseases PCR Polymerase Chain Reaction - methods Pregnancy Pregnancy. Fetus. Placenta prenatal diagnosis Prenatal Diagnosis - methods Protozoal diseases Spectrometry, Fluorescence Toxoplasma - genetics Toxoplasma - immunology Toxoplasmosis Toxoplasmosis, Congenital - diagnosis Ultrasonography, Prenatal |
| title | Prenatal diagnosis of congenital toxoplasmosis by duplex real-time PCR using fluorescence resonance energy transfer hybridization probes |
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