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Multiplex primer prediction software for divergent targets

Identifieur interne : 000712 ( Ncbi/Merge ); précédent : 000711; suivant : 000713

Multiplex primer prediction software for divergent targets

Auteurs : Shea N. Gardner ; Amy L. Hiddessen ; Peter L. Williams ; Christine Hara ; Mark C. Wagner ; Bill W. Colston

Source :

RBID : PMC:2770652

Descripteurs français

English descriptors

Abstract

Abstract

We describe a Multiplex Primer Prediction (MPP) algorithm to build multiplex compatible primer sets to amplify all members of large, diverse and unalignable sets of target sequences. The MPP algorithm is scalable to larger target sets than other available software, and it does not require a multiple sequence alignment. We applied it to questions in viral detection, and demonstrated that there are no universally conserved priming sequences among viruses and that it could require an unfeasibly large number of primers (∼3700 18-mers or ∼2000 10-mers) to generate amplicons from all sequenced viruses. We then designed primer sets separately for each viral family, and for several diverse species such as foot-and-mouth disease virus (FMDV), hemagglutinin (HA) and neuraminidase (NA) segments of influenza A virus, Norwalk virus, and HIV-1. We empirically demonstrated the application of the software with a multiplex set of 16 short (10 nt) primers designed to amplify the Poxviridae family to produce a specific amplicon from vaccinia virus.


Url:
DOI: 10.1093/nar/gkp659
PubMed: 19759213
PubMed Central: 2770652

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PMC:2770652

Le document en format XML

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<front>
<div type="abstract" xml:lang="en">
<title>Abstract</title>
<p>We describe a Multiplex Primer Prediction (MPP) algorithm to build multiplex compatible primer sets to amplify all members of large, diverse and unalignable sets of target sequences. The MPP algorithm is scalable to larger target sets than other available software, and it does not require a multiple sequence alignment. We applied it to questions in viral detection, and demonstrated that there are no universally conserved priming sequences among viruses and that it could require an unfeasibly large number of primers (∼3700 18-mers or ∼2000 10-mers) to generate amplicons from all sequenced viruses. We then designed primer sets separately for each viral family, and for several diverse species such as foot-and-mouth disease virus (FMDV), hemagglutinin (HA) and neuraminidase (NA) segments of influenza A virus, Norwalk virus, and HIV-1. We empirically demonstrated the application of the software with a multiplex set of 16 short (10 nt) primers designed to amplify the Poxviridae family to produce a specific amplicon from vaccinia virus.</p>
</div>
</front>
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<term>Algorithms</term>
<term>DNA (analysis)</term>
<term>DNA Primers (chemistry)</term>
<term>Humans</term>
<term>RNA Viruses (genetics)</term>
<term>Sequence Analysis, DNA</term>
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<term>Vaccinia virus (genetics)</term>
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<term>ADN (analyse)</term>
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<term>Amorces ADN ()</term>
<term>Analyse de séquence d'ADN</term>
<term>Humains</term>
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<term>Virus (isolement et purification)</term>
<term>Virus de la vaccine (génétique)</term>
<term>Virus à ARN (génétique)</term>
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<term>DNA</term>
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<term>DNA Primers</term>
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<term>ADN</term>
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<term>RNA Viruses</term>
<term>Vaccinia virus</term>
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<term>Algorithms</term>
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<front>
<div type="abstract" xml:lang="en">We describe a Multiplex Primer Prediction (MPP) algorithm to build multiplex compatible primer sets to amplify all members of large, diverse and unalignable sets of target sequences. The MPP algorithm is scalable to larger target sets than other available software, and it does not require a multiple sequence alignment. We applied it to questions in viral detection, and demonstrated that there are no universally conserved priming sequences among viruses and that it could require an unfeasibly large number of primers ( approximately 3700 18-mers or approximately 2000 10-mers) to generate amplicons from all sequenced viruses. We then designed primer sets separately for each viral family, and for several diverse species such as foot-and-mouth disease virus (FMDV), hemagglutinin (HA) and neuraminidase (NA) segments of influenza A virus, Norwalk virus, and HIV-1. We empirically demonstrated the application of the software with a multiplex set of 16 short (10 nt) primers designed to amplify the Poxviridae family to produce a specific amplicon from vaccinia virus.</div>
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