Multiplex detection and SNP genotyping in a single fluorescence channel.
Probe-based PCR is widely used for SNP (single nucleotide polymorphism) genotyping and pathogen nucleic acid detection due to its simplicity, sensitivity and cost-effectiveness. However, the multiplex capability of hydrolysis probe-based PCR is normally limited to one target (pathogen or allele) per...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2012-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3260291?pdf=render |
| _version_ | 1828848991015534592 |
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| author | Guoliang Fu Andrea Miles Luke Alphey |
| author_facet | Guoliang Fu Andrea Miles Luke Alphey |
| author_sort | Guoliang Fu |
| collection | DOAJ |
| description | Probe-based PCR is widely used for SNP (single nucleotide polymorphism) genotyping and pathogen nucleic acid detection due to its simplicity, sensitivity and cost-effectiveness. However, the multiplex capability of hydrolysis probe-based PCR is normally limited to one target (pathogen or allele) per fluorescence channel. Current fluorescence PCR machines typically have 4-6 channels. We present a strategy permitting the multiplex detection of multiple targets in a single detection channel. The technique is named Multiplex Probe Amplification (MPA). Polymorphisms of the CYP2C9 gene (cytochrome P450, family 2, subfamily C, polypeptide 9, CYP2C9*2) and human papillomavirus sequences HPV16, 18, 31, 52 and 59 were chosen as model targets for testing MPA. The allele status of the CYP2C9*2 determined by MPA was entirely concordant with the reference TaqMan® SNP Genotyping Assays. The four HPV strain sequences could be independently detected in a single fluorescence detection channel. The results validate the multiplex capacity, the simplicity and accuracy of MPA for SNP genotyping and multiplex detection using different probes labeled with the same fluorophore. The technique offers a new way to multiplex in a single detection channel of a closed-tube PCR. |
| first_indexed | 2024-12-12T22:42:49Z |
| format | Article |
| id | doaj.art-e418430092c645eca66fe213a464b023 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-12T22:42:49Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-e418430092c645eca66fe213a464b0232022-12-22T00:09:18ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0171e3034010.1371/journal.pone.0030340Multiplex detection and SNP genotyping in a single fluorescence channel.Guoliang FuAndrea MilesLuke AlpheyProbe-based PCR is widely used for SNP (single nucleotide polymorphism) genotyping and pathogen nucleic acid detection due to its simplicity, sensitivity and cost-effectiveness. However, the multiplex capability of hydrolysis probe-based PCR is normally limited to one target (pathogen or allele) per fluorescence channel. Current fluorescence PCR machines typically have 4-6 channels. We present a strategy permitting the multiplex detection of multiple targets in a single detection channel. The technique is named Multiplex Probe Amplification (MPA). Polymorphisms of the CYP2C9 gene (cytochrome P450, family 2, subfamily C, polypeptide 9, CYP2C9*2) and human papillomavirus sequences HPV16, 18, 31, 52 and 59 were chosen as model targets for testing MPA. The allele status of the CYP2C9*2 determined by MPA was entirely concordant with the reference TaqMan® SNP Genotyping Assays. The four HPV strain sequences could be independently detected in a single fluorescence detection channel. The results validate the multiplex capacity, the simplicity and accuracy of MPA for SNP genotyping and multiplex detection using different probes labeled with the same fluorophore. The technique offers a new way to multiplex in a single detection channel of a closed-tube PCR.http://europepmc.org/articles/PMC3260291?pdf=render |
| spellingShingle | Guoliang Fu Andrea Miles Luke Alphey Multiplex detection and SNP genotyping in a single fluorescence channel. PLoS ONE |
| title | Multiplex detection and SNP genotyping in a single fluorescence channel. |
| title_full | Multiplex detection and SNP genotyping in a single fluorescence channel. |
| title_fullStr | Multiplex detection and SNP genotyping in a single fluorescence channel. |
| title_full_unstemmed | Multiplex detection and SNP genotyping in a single fluorescence channel. |
| title_short | Multiplex detection and SNP genotyping in a single fluorescence channel. |
| title_sort | multiplex detection and snp genotyping in a single fluorescence channel |
| url | http://europepmc.org/articles/PMC3260291?pdf=render |
| work_keys_str_mv | AT guoliangfu multiplexdetectionandsnpgenotypinginasinglefluorescencechannel AT andreamiles multiplexdetectionandsnpgenotypinginasinglefluorescencechannel AT lukealphey multiplexdetectionandsnpgenotypinginasinglefluorescencechannel |