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Discriminatory power of RNA family models.

ABSTRACT: MOTIVATION: RNA family models group nucleotide sequences that share a common biological function. These models can be used to find new sequences belonging to the same family. To succeed in this task, a model needs to exhibit high sensitivity as well as high specificity. As model construction is guided by a manual process, a number of problems can occur, such as the introduction of more than one model for the same family or poorly constructed models. We explore the Rfam database to discover such problems. RESULTS: Our main contribution is in the definition of the discriminatory power of RNA family models, together with a first algorithm for its computation. In addition, we present calculations across the whole Rfam database that show several families lacking high specificity when compared to other families. We give a list of these clusters of families and provide a tentative explanation. Our program can be used to: (i) make sure that new models are not equivalent to any model already present in the database; and (ii) new models are not simply submodels of existing families. AVAILABILITY: www.tbi.univie.ac.at/software/cmcompare/. The code is licensed under the GPLv3. Results for the whole Rfam database and supporting scripts are available together with the software.

SUBMITTER: Honer zu Siederdissen C

PROVIDER: S-EPMC2935435 | biostudies-literature | 2010 Sep

REPOSITORIES: biostudies-literature

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Discriminatory power of RNA family models.

Höner zu Siederdissen Christian C Hofacker Ivo L IL

Bioinformatics (Oxford, England) 20100901 18

<h4>Motivation</h4>RNA family models group nucleotide sequences that share a common biological function. These models can be used to find new sequences belonging to the same family. To succeed in this task, a model needs to exhibit high sensitivity as well as high specificity. As model construction is guided by a manual process, a number of problems can occur, such as the introduction of more than one model for the same family or poorly constructed models. We explore the Rfam database to discove ...[more]

PMID: 20823307

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Project description:BackgroundSeveral technical hurdles and limitations have restricted the use of IS6110 restriction fragment length polymorphism (IS6110 RFLP), the most effective typing method for detecting recent tuberculosis (TB) transmission events. This has prompted us to conceive an alternative modality, IS6110-5'3'FP, a plasmid-based cloning approach coupled to a single PCR amplification of differentially labeled 5' and 3' IS6110 polymorphic ends and their automated fractionation on a capillary sequencer. The potential of IS6110-5'3'FP to be used as an alternative to IS6110 RFLP has been previously demonstrated, yet further technical improvements are still required for optimal discriminatory power and versatility.ObjectivesHere we introduced critical amendments to the original IS6110-5'3'FP protocol and compared its performance to that of 24-loci multiple interspersed repetitive unit-variable number tandem repeats (MIRU-VNTR), the current standard method for TB transmission analyses.MethodsIS6110-5'3'FP protocol modifications involved: (i) the generation of smaller-sized polymorphic fragments for efficient cloning and PCR amplification, (ii) omission of the plasmid amplification step in E. coli for shorter turnaround times, (iii) the use of more stable fluorophores for increased sensitivity, (iv) automated subtraction of background fluorescent signals, and (v) the automated conversion of fluorescent peaks into binary data.ResultsIn doing so, the overall turnaround time of IS6110-5'3'FP was reduced to 4 hours. The new protocol allowed detecting almost all 5' and 3' IS6110 polymorphic fragments of any given strain, including IS6110 high-copy number Beijing strains. IS6110-5'3'FP proved much more discriminative than 24-loci MIRU-VNTR, particularly with strains of the M. tuberculosis lineage 4.ConclusionsThe IS6110-5'3'FP protocol described herein reached the optimal discriminatory potential of IS6110 fingerprinting and proved more accurate than 24-loci MIRU-VNTR in estimating recent TB transmission. The method, which is highly cost-effective, was rendered versatile enough to prompt its evaluation as an automatized solution for a TB integrated molecular surveillance.

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Discriminatory power of RNA family models.

Publications

Discriminatory power of RNA family models.

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