NuMY—A qPCR Assay Simultaneously Targeting Human Autosomal, Y-Chromosomal, and Mitochondrial DNA
The accurate quantification of DNA in forensic samples is of utmost importance. These samples are often present in limited amounts; therefore, it is indicated to use the appropriate analysis route with the optimum DNA amount (when possible). Also, DNA quantification can inform about the degradation...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-08-01
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| Series: | Genes |
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| Online Access: | https://www.mdpi.com/2073-4425/14/8/1645 |
| _version_ | 1797584643904503808 |
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| author | Catarina Xavier Charlotte Sutter Christina Amory Harald Niederstätter Walther Parson |
| author_facet | Catarina Xavier Charlotte Sutter Christina Amory Harald Niederstätter Walther Parson |
| author_sort | Catarina Xavier |
| collection | DOAJ |
| description | The accurate quantification of DNA in forensic samples is of utmost importance. These samples are often present in limited amounts; therefore, it is indicated to use the appropriate analysis route with the optimum DNA amount (when possible). Also, DNA quantification can inform about the degradation stage and therefore support the decision on which downstream genotyping method to use. Consequently, DNA quantification aids in getting the best possible results from a forensic sample, considering both its DNA quantity and quality limitations. Here, we introduce NuMY, a new quantitative real-time PCR (qPCR) method for the parallel quantification of human nuclear (n) and mitochondrial (mt) DNA, assessing the male portion in mixtures of both sexes and testing for possible PCR inhibition. NuMY is based on previous work and follows the MIQE guidelines whenever applicable. Although quantification of nuclear (n)DNA by simultaneously analyzing autosomal and male-specific targets is available in commercial qPCR kits, tools that include the quantification of mtDNA are sparse. The quantification of mtDNA has proven relevant for samples with low nDNA content when conventional DNA fingerprinting techniques cannot be followed. Furthermore, the development and use of new massively parallel sequencing assays that combine multiple marker types, i.e., autosomal, Y-chromosomal, and mtDNA, can be optimized when precisely knowing the amount of each DNA component present in the input sample. For high-quality DNA extracts, NuMY provided nDNA results comparable to those of another quantification technique and has also proven to be a reliable tool for challenging, forensically relevant samples such as mixtures, inhibited, and naturally degraded samples. |
| first_indexed | 2024-03-10T23:54:36Z |
| format | Article |
| id | doaj.art-ae0e24c9ca0c446c8c9030bb4c33a9a6 |
| institution | Directory Open Access Journal |
| issn | 2073-4425 |
| language | English |
| last_indexed | 2024-03-10T23:54:36Z |
| publishDate | 2023-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Genes |
| spelling | doaj.art-ae0e24c9ca0c446c8c9030bb4c33a9a62023-11-19T01:16:26ZengMDPI AGGenes2073-44252023-08-01148164510.3390/genes14081645NuMY—A qPCR Assay Simultaneously Targeting Human Autosomal, Y-Chromosomal, and Mitochondrial DNACatarina Xavier0Charlotte Sutter1Christina Amory2Harald Niederstätter3Walther Parson4Institute of Legal Medicine, Medical University of Innsbruck, 6020 Innsbruck, AustriaInstitute of Legal Medicine, Medical University of Innsbruck, 6020 Innsbruck, AustriaInstitute of Legal Medicine, Medical University of Innsbruck, 6020 Innsbruck, AustriaInstitute of Legal Medicine, Medical University of Innsbruck, 6020 Innsbruck, AustriaInstitute of Legal Medicine, Medical University of Innsbruck, 6020 Innsbruck, AustriaThe accurate quantification of DNA in forensic samples is of utmost importance. These samples are often present in limited amounts; therefore, it is indicated to use the appropriate analysis route with the optimum DNA amount (when possible). Also, DNA quantification can inform about the degradation stage and therefore support the decision on which downstream genotyping method to use. Consequently, DNA quantification aids in getting the best possible results from a forensic sample, considering both its DNA quantity and quality limitations. Here, we introduce NuMY, a new quantitative real-time PCR (qPCR) method for the parallel quantification of human nuclear (n) and mitochondrial (mt) DNA, assessing the male portion in mixtures of both sexes and testing for possible PCR inhibition. NuMY is based on previous work and follows the MIQE guidelines whenever applicable. Although quantification of nuclear (n)DNA by simultaneously analyzing autosomal and male-specific targets is available in commercial qPCR kits, tools that include the quantification of mtDNA are sparse. The quantification of mtDNA has proven relevant for samples with low nDNA content when conventional DNA fingerprinting techniques cannot be followed. Furthermore, the development and use of new massively parallel sequencing assays that combine multiple marker types, i.e., autosomal, Y-chromosomal, and mtDNA, can be optimized when precisely knowing the amount of each DNA component present in the input sample. For high-quality DNA extracts, NuMY provided nDNA results comparable to those of another quantification technique and has also proven to be a reliable tool for challenging, forensically relevant samples such as mixtures, inhibited, and naturally degraded samples.https://www.mdpi.com/2073-4425/14/8/1645DNA quantificationhuman nuclear DNAY chromosomemitochondrial DNA |
| spellingShingle | Catarina Xavier Charlotte Sutter Christina Amory Harald Niederstätter Walther Parson NuMY—A qPCR Assay Simultaneously Targeting Human Autosomal, Y-Chromosomal, and Mitochondrial DNA Genes DNA quantification human nuclear DNA Y chromosome mitochondrial DNA |
| title | NuMY—A qPCR Assay Simultaneously Targeting Human Autosomal, Y-Chromosomal, and Mitochondrial DNA |
| title_full | NuMY—A qPCR Assay Simultaneously Targeting Human Autosomal, Y-Chromosomal, and Mitochondrial DNA |
| title_fullStr | NuMY—A qPCR Assay Simultaneously Targeting Human Autosomal, Y-Chromosomal, and Mitochondrial DNA |
| title_full_unstemmed | NuMY—A qPCR Assay Simultaneously Targeting Human Autosomal, Y-Chromosomal, and Mitochondrial DNA |
| title_short | NuMY—A qPCR Assay Simultaneously Targeting Human Autosomal, Y-Chromosomal, and Mitochondrial DNA |
| title_sort | numy a qpcr assay simultaneously targeting human autosomal y chromosomal and mitochondrial dna |
| topic | DNA quantification human nuclear DNA Y chromosome mitochondrial DNA |
| url | https://www.mdpi.com/2073-4425/14/8/1645 |
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