A novel SYBR-based duplex qPCR for the detection of gene dosage: detection of an <it>APC</it> large deletion in a familial adenomatous polyposis patient with an unusual phenotype
<p>Abstract</p> <p>Background</p> <p>Familial adenomatous polyposis (FAP) is a hereditary colorectal cancer syndrome caused by a loss of function of the <it>APC</it> gene. Large deletions in <it>APC</it> are a common cause of FAP; despite the exi...
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| Format: | Article |
| Language: | English |
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BMC
2012-07-01
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| Series: | BMC Medical Genetics |
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| Online Access: | http://www.biomedcentral.com/1471-2350/13/55 |
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| author | Torrezan Giovana da Silva Felipe Cavalcanti Krepischi Ana Cristina dos Santos Érika Maria Rossi Benedito Carraro Dirce |
| author_facet | Torrezan Giovana da Silva Felipe Cavalcanti Krepischi Ana Cristina dos Santos Érika Maria Rossi Benedito Carraro Dirce |
| author_sort | Torrezan Giovana |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Familial adenomatous polyposis (FAP) is a hereditary colorectal cancer syndrome caused by a loss of function of the <it>APC</it> gene. Large deletions in <it>APC</it> are a common cause of FAP; despite the existence of a variety of gene dosage detection methodologies, most are labor intensive and time and resource consuming.</p> <p>Methods</p> <p>We describe a new duplex qPCR method for gene dosage analysis based on the coamplification of a target and a reference gene in a SYBR Green reaction, followed by a comparison of the ratio between the target and the reference peaks of the melting curve for the test (patient) and control samples. The reliability of the described duplex qPCR was validated for several genes (<it>APC</it>, <it>HPRT1</it>, <it>ATM</it>, <it>PTEN</it> and <it>BRCA1</it>).</p> <p>Results</p> <p>Using this novel gene dosage method, we have identified an <it>APC</it> gene deletion in a FAP patient undergoing genetic testing. Comparative genomic hybridization based on microarrays (aCGH) was used to confirm and map the extent of the deletion, revealing a 5.2 MB rearrangement (5q21.3-q22.3) encompassing the entire <it>APC</it> and 19 additional genes.</p> <p>Conclusion</p> <p>The novel assay accurately detected losses and gains of one copy of the target sequences, representing a reliable and flexible alternative to other gene dosage techniques. In addition, we described a FAP patient harboring a gross deletion at 5q21.3-q22.3 with an unusual phenotype of the absence of mental impairment and dysmorphic features.</p> |
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| institution | Directory Open Access Journal |
| issn | 1471-2350 |
| language | English |
| last_indexed | 2024-12-19T14:42:02Z |
| publishDate | 2012-07-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Medical Genetics |
| spelling | doaj.art-6c803dc472c84baf80c82ceae093f2c42022-12-21T20:17:03ZengBMCBMC Medical Genetics1471-23502012-07-011315510.1186/1471-2350-13-55A novel SYBR-based duplex qPCR for the detection of gene dosage: detection of an <it>APC</it> large deletion in a familial adenomatous polyposis patient with an unusual phenotypeTorrezan Giovanada Silva Felipe CavalcantiKrepischi Ana Cristinados Santos Érika MariaRossi BeneditoCarraro Dirce<p>Abstract</p> <p>Background</p> <p>Familial adenomatous polyposis (FAP) is a hereditary colorectal cancer syndrome caused by a loss of function of the <it>APC</it> gene. Large deletions in <it>APC</it> are a common cause of FAP; despite the existence of a variety of gene dosage detection methodologies, most are labor intensive and time and resource consuming.</p> <p>Methods</p> <p>We describe a new duplex qPCR method for gene dosage analysis based on the coamplification of a target and a reference gene in a SYBR Green reaction, followed by a comparison of the ratio between the target and the reference peaks of the melting curve for the test (patient) and control samples. The reliability of the described duplex qPCR was validated for several genes (<it>APC</it>, <it>HPRT1</it>, <it>ATM</it>, <it>PTEN</it> and <it>BRCA1</it>).</p> <p>Results</p> <p>Using this novel gene dosage method, we have identified an <it>APC</it> gene deletion in a FAP patient undergoing genetic testing. Comparative genomic hybridization based on microarrays (aCGH) was used to confirm and map the extent of the deletion, revealing a 5.2 MB rearrangement (5q21.3-q22.3) encompassing the entire <it>APC</it> and 19 additional genes.</p> <p>Conclusion</p> <p>The novel assay accurately detected losses and gains of one copy of the target sequences, representing a reliable and flexible alternative to other gene dosage techniques. In addition, we described a FAP patient harboring a gross deletion at 5q21.3-q22.3 with an unusual phenotype of the absence of mental impairment and dysmorphic features.</p>http://www.biomedcentral.com/1471-2350/13/55Gene dosageQuantitative PCRFamilial adenomatous polyposis<it>APC</it> whole gene deletion |
| spellingShingle | Torrezan Giovana da Silva Felipe Cavalcanti Krepischi Ana Cristina dos Santos Érika Maria Rossi Benedito Carraro Dirce A novel SYBR-based duplex qPCR for the detection of gene dosage: detection of an <it>APC</it> large deletion in a familial adenomatous polyposis patient with an unusual phenotype BMC Medical Genetics Gene dosage Quantitative PCR Familial adenomatous polyposis <it>APC</it> whole gene deletion |
| title | A novel SYBR-based duplex qPCR for the detection of gene dosage: detection of an <it>APC</it> large deletion in a familial adenomatous polyposis patient with an unusual phenotype |
| title_full | A novel SYBR-based duplex qPCR for the detection of gene dosage: detection of an <it>APC</it> large deletion in a familial adenomatous polyposis patient with an unusual phenotype |
| title_fullStr | A novel SYBR-based duplex qPCR for the detection of gene dosage: detection of an <it>APC</it> large deletion in a familial adenomatous polyposis patient with an unusual phenotype |
| title_full_unstemmed | A novel SYBR-based duplex qPCR for the detection of gene dosage: detection of an <it>APC</it> large deletion in a familial adenomatous polyposis patient with an unusual phenotype |
| title_short | A novel SYBR-based duplex qPCR for the detection of gene dosage: detection of an <it>APC</it> large deletion in a familial adenomatous polyposis patient with an unusual phenotype |
| title_sort | novel sybr based duplex qpcr for the detection of gene dosage detection of an it apc it large deletion in a familial adenomatous polyposis patient with an unusual phenotype |
| topic | Gene dosage Quantitative PCR Familial adenomatous polyposis <it>APC</it> whole gene deletion |
| url | http://www.biomedcentral.com/1471-2350/13/55 |
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