Title |
Optimizing multiplex SNP-based data analysis for genotyping of Mycobacterium tuberculosis isolates
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Published in |
BMC Genomics, July 2014
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DOI | 10.1186/1471-2164-15-572 |
Pubmed ID | |
Authors |
Sarah Sengstake, Nino Bablishvili, Anja Schuitema, Nino Bzekalava, Edgar Abadia, Jessica de Beer, Nona Tadumadze, Maka Akhalaia, Kiki Tuin, Nestani Tukvadze, Rusudan Aspindzelashvili, Elizabeta Bachiyska, Stefan Panaiotov, Christophe Sola, Dick van Soolingen, Paul Klatser, Richard Anthony, Indra Bergval |
Abstract |
Multiplex ligation-dependent probe amplification (MLPA) is a powerful tool to identify genomic polymorphisms. We have previously developed a single nucleotide polymorphism (SNP) and large sequence polymorphisms (LSP)-based MLPA assay using a read out on a liquid bead array to screen for 47 genetic markers in the Mycobacterium tuberculosis genome. In our assay we obtain information regarding the Mycobacterium tuberculosis lineage and drug resistance simultaneously. Previously we called the presence or absence of a genotypic marker based on a threshold signal level. Here we present a more elaborate data analysis method to standardize and streamline the interpretation of data generated by MLPA. The new data analysis method also identifies intermediate signals in addition to classification of signals as positive and negative. Intermediate calls can be informative with respect to identifying the simultaneous presence of sensitive and resistant alleles or infection with multiple different Mycobacterium tuberculosis strains. |
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