Background: Left ventricular noncompaction (LVNC) is a myocardial abnormality char-acterized by prominent trabeculations, a thin compacted layer, and deep intertrabecular recesses. LVNC is a heterogeneous myocardial disorder of uncertain etiology, which may occur as a congenital or acquired condition or develop in association with other cardio-myopathies. Genetic factors have also been implicated in its pathogenesis. Objectives: The aim of this study was to investigate the genetic background of patients with LVNC through next-generation sequencing (NGS) analysis and to identify potentially dis-ease-associated variants. Methods: Twenty-five patients with LVNC underwent genetic testing by NGS using the Ion Torrent™ platform. Variants identified by NGS were con-firmed by Sanger sequencing. The interpretation was performed according to ACMG guidelines using multiple bioinformatic and clinical databases. Results: Genetic variants were identified in 13 patients, with 14 variants detected in 9 genes (TTN, MYH7, CTNNA3, DSG2, FLNC, JPH2, NKX2-6, TNNC1, and JUP). Eight variants (57%) were nov-el, four (29%) had been previously reported in clinical cases without scientific discussion, and two (14%) were already described in the literature. According to ACMG criteria, vari-ants were classified as pathogenic (n=1), likely pathogenic (n=7), or variants of uncertain significance (n=5). Conclusions: These findings highlight the marked genetic heterogenei-ty of LVNC and support the role of genetic testing in improving diagnosis, risk stratifica-tion and clinical management.

Background/Objectives: SUDEP is the sudden, unexpected death of someone with epilepsy, and occurs mainly during sleep or at rest, or when the individual does not seem to have experienced a convulsive seizure. The cause of death in SUDEP is still unknown, and it may differ between cases. Cardiac factors are among the most prevalent causes observed in SUDEP. Therefore, within the forensic medicine framework, identifying well-known DNA markers involved in cardiac sudden and unexpected death would aid in understanding the cause of SUDEP, as well as in finding cardiac risk markers in patients with epilepsy. The purpose of this study was to identify any genetic variants by analyzing blood and formalin-fixed paraffin-embedded (FFPE) tissue samples, utilizing next-generation sequencing techniques. Methods: We investigated five cases of SUDEP that were examined at the Legal Medicine department of Ancona (Italy). Peripheral blood or FFPE cardiac tissues were collected, and different DNA extraction methods were performed. In particular, this study underlines a new extraction method from FFPE tissue, adapting the Casework kit for forensic application to our purpose. Later, about one hundred genes correlated to inherited cardiac diseases were sequenced through the Ion PGM System and Ion GeneStudio S5 Systems. Results: Bioinformatic analysis showed some genetic variants of unknown significance (VUS) on genes involved in SUDEP: RYR2, SCN8A, and AKAP9. Conclusions: As expected, very low coverage of the target base was observed for FFPE tissue samples because of the complexity of the biological material. Therefore, the presence of any significant variants in unamplified regions cannot be excluded in the FFPE samples. As suggested by the literature, the variants found in the blood samples are potentially associated with SUDEP.

Background/Objectives: The study of DNA transfer and persistence has become increasingly significant, driven by advancements in DNA detection sensitivity and the need for reliable forensic evidence. In forensic investigations, saliva and saliva-stained materials are recognised as valuable DNA sources, particularly in cases of homicide, sexual assault, and burglary, where saliva can be transferred between individuals during the criminal act. The time between the crime and sample collection is a critical factor that can influence the success of the analysis. The value of the specimens collected from the victim’s skin or mouth (perilabial and labial sites, teeth and tongue) after the crime has not been investigated with currently used highly sensitive and specific molecular methods. Methods: On the assumption that a significant loss of DNA occurred, in our study, 10 voluntary pairs were tested at different time points after intense kissing and samples were taken from the above-mentioned sites to assess the presence of the donor’s DNA. Extracted DNA was quantified using the Plexor HY System kit (Promega), and both autosomal STRs and Y-STRs were analysed. Results: The results reveal a greater persistence of male DNA on the female partner, particularly in the labial and perilabial regions, even up to 120 min after contact, in terms of both concentration and duration. Conclusions: This study emphasises the forensic importance of salivary DNA as a solid source of evidence, particularly in investigations involving mixed DNA profiles.

Quantification of human DNA is key in forensic genetics. A more accurate estimate of the amount of DNA is essential for planning and optimising genotyping assays, as is evaluating the presence of PCR inhibitory substances and DNA degradation status. Multiplex qPCR assays are helpful in forensics because they can quantify different targets simultaneously, thus saving valuable samples, time, and labour. The aim of this study was to highlight the challenges in the developmental validation of a multiplex real-time PCR assay and the drawbacks encountered in translating a previously described and validated assay (SD quants) to a different technology by modifying the dye probes and reagent mix to be used in a different instrument. We developed a TaqMan probe-based multiplex qPCR using reagents and fluorescent probes adapted for the Rotor-Gene 6000 instrument (QIAGEN, Hilden, Germany). The initial assay combined two mitochondrial DNA (mtDNA) and two nuclear DNA (nDNA) targets, with amplification products of different sizes (mtDNA = 69 and 143 bp; nDNA = 71 and 181 bp), to estimate the DNA degradation status and an internal positive control (IPC) to detect potential inhibitors. During the initial testing of the assay, we observed an interaction between the 69 bp mtDNA target and the 71 bp nDNA target probe, and experiments were conducted to resolve this issue without success. We removed the small nDNA target (71 bp) and changed from a 5-plex to a 4-plex qPCR assay (qMIND). The final tetraplex assay was tested on 105 forensic samples and/or small amounts of degraded DNA, such as bones, teeth, fingernails, formalin-fixed paraffin-embedded tissues (FFPE), and hair shaft samples. The quantification results were compared with data acquired from the same samples using another commercially available quantification system commonly used in forensic laboratories. In addition, the short tandem repeat (STR) profiles were investigated to determine their correlation with the quantitative values obtained. Overall, the qPCR assay was robust and reliable for DNA quantification in samples commonly used in forensic practice.

Background and objectives: Sudden cardiac death (SCD) is a natural and unexpected death of cardiac origin that occurs within 1 h from the onset of acute symptoms. The major leading causes of SCD are cardiomyopathies and channelopathies. In this review, we focus on channelopathies, inherited diseases caused by mutations affecting genes encoding membrane ion channels (sodium, potassium or calcium channels) or cellular structures that affect Ca2+ availability. The diagnosis of diseases such as long QT, Brugada syndrome, short QT and catecholaminergic polymorphic ventricular tachycardia (CPVT) is still challenging. Currently, genetic testing and next-generation sequencing allow us to identify many rare alterations. However, some non-coding variants, e.g., splice-site variants, are usually difficult to interpret and to classify. Methods: In our review, we searched for splice-site variants of genes involved in channelopathies, focusing on variants of unknown significance (VUSs) registered on ClinVar up to now. Results: The research led to a high number of splice-site VUSs of genes involved in channelopathies, suggesting the performance of deeper studies. Conclusions: In order to interpret the correlation between variants and pathologies, we discuss experimental studies, such as RNA sequencing and functional analysis of proteins. Unfortunately, as these in vitro analyses cannot always be performed, we draw attention to in silico studies as future perspectives in genetics. This review has the aim of discussing the potential methods of detection and interpretation of VUSs, bringing out the need for a future reclassification of variants with currently unknown significance.

The introduction of Massive Parallel Sequencing technology (MPS) in the forensic genetics field has opened new possibilities in forensic DNA genotyping. The advantage of MPS is multifold, including: high throughput sequencing; production of millions of DNA molecules in parallel; simultaneous analysis of large number of markers; as well as different type of markers; and a high number of sample in a single experimental run. Beside genotyping traditional forensic markers for identification, i.e., Short Tandem Repeat (STR), Single Nucleotide Polymorphism (SNP), and mitochondrial DNA (mtDNA), MPS offers the potential to genotype a new type of genetic marker, known as microhaplotypes. Moreover, MPS makes it possible to explore the potential of forensic DNA phenotyping and of the forensic transcriptomic. This article discusses different MPS approaches used in forensic, the applications in forensic field, and benefits and drawbacks are discussed.

Abstract Background and Methods The management of patients with Cardiac Amyloidosis (CA) and Hypertrophic Cardiomyopathy (HCM) is complex and requires specific skills. Collaboration between centres remains essential because not all centres can perform complex diagnostic techniques (such as endomyocardial biopsy and mass spectrometry) or prescribe disease–modifying therapies, Therefore, in 2019 we created a collaborative network for diagnosis and treatment of CA (TTR and AL) and HCM (sarcomeric and phenocopies) with the aim of enhancing disease awareness among physicians and favoring appropriate access to innovative diagnostic tools/therapies. The network includes all 14 Cardiology Departments and outpatient clinics from 5 Local Health Units. At Referral Centre of the Regional University Hospital, all diagnostic and therapeutic tools necessary for a comprehensive management of CM patients including genetic testing, endomyocardial biopsy, ablation of complex ventricular arrhythmias, and septal myectomy for HOCM, are currently available and at disposal for the entire network. Results Thanks to this network, from December 2019 to December 2022, 283 patients were referred to our Center (fig. 1). Of these, 136 (48%) had CA, 122 (43%) were diagnosed with HCM (68 with obstructive form and 54 with non–obstructive form), 12 (4%) with Anderson Fabry Disease (AFD) and 13 (5%) with other forms of cardiomyopathy (miscellanea: arrhythmogenic, LVNC, etc). As for temporal trends in diagnosis of CM patients, in the first 2 years the diagnosis of HCM was more frequent than that of CA, while the number of CA patients has steadily grown, overtaking that of HCM patients in the last year (fig.2). The number of AFD diagnoses was consistently low over the 3 years, despite the spread use of genetic testing in all HCM patients. All patients with CA (136) were managed according to current algorythm with need of EMB in uncertain cases (8). Of the 136 patients with AC, 122 had TTR while 14 had AL (Fig.3). All patients with ATTR (122) underwent genetic testing: in 14 cases the presence of mutations in the TTR gene was documented (8 Ile68Leu; 4 Val30Met; 2 Val122Ile). 51 ATTR patients were treated with tafamidis, while 3 patients with variant–TTR and neuropathy received patisiran iv. Conclusions Implementing specific clinical network provided excellent results, allowing a precise phenotype/genotype characterization and favoring appropriate access to specific disease–modifying drugs.

Sudden cardiac death (SCD) is one of the leading causes of death in the world and for this reason it has attracted the attention of numerous researchers in the field of legal medicine. It is not easy to determine the cause in a SCD case and the available methods used for diagnosis cannot always give an exhaustive answer. In addition, the molecular analysis of genes does not lead to a clear conclusion, but it could be interesting to focus attention on the expression level of miRNAs, a class of non-coding RNA of about 22 nucleotides. The role of miRNAs is to regulate the gene expression through complementary binding to 3′-untraslated regions of miRNAs, leading to the inhibition of translation or to mRNA degradation. In recent years, several studies were performed with the aim of exploring the use of these molecules as biomarkers for SCD cases, and to also distinguish the causes that lead to cardiac death. In this review, we summarize experiments, evidence, and results of different studies on the implication of miRNAs in SCD cases. We discuss the different biological starting materials with their respective advantages and disadvantages, studying miRNA expression on tissue (fresh-frozen tissue and FFPE tissue), circulating cell-free miRNAs in blood of patients affected by cardiac disease at high risk of SCD, and exosomal miRNAs analyzed from serum of people who died from SCD.

The growing use of massively parallel sequencing (MPS) for the whole mitochondrial genome analysis in forensic laboratories, requires the establishment of efficient workflows and interpretation procedures, to support the feasibility of the technology and the reliability of the data. In the case of reference samples, such as blood and buccal swabs, the generation of mtDNA profiles by using MPS is relatively simple. Conversely, many forensic casework samples still pose challenges for the MPS, data interpretation and reporting of mtDNA. This is especially true for the analysis of shed hairs, which are one of the most common evidence types and which are among the most limited in terms of DNA quantity and quality. Due to these limitations, every step involved in the analysis become essentials and should be performed in order to obtain the best performance, optimizing the outcomes and minimizing the errors. In light of this, we present a study focusing on the extraction, quantification and MPS of the whole mtDNA in hair shafts, with the aims of set-up and validate a methodological pipeline to obtain the best sequencing results. The overall performance of the MPS panel, mainly in terms of total coverage, amplicons coverage and different primer pools efficiency, was evaluated also in relation to the different hair fragments, the mtDNA copy number used for libraries preparation and its degradation state.

The performance of the Precision ID Identity Panel (Thermo Fisher Scientific) was assessed on a set of 87 forensic samples with different levels of degradation for which a reference sample from the "same donor" or from a "first degree relative" was available. PCR-MPS analysis was performed with DNA input ranging from 1 ng to 12 pg and through 21-26 PCR cycles, in replicate tests, and a total number of 255 libraries were sequenced on the Ion Personal Genome Machine™ (PGM™) System. The evaluation of the molecular data allowed to set a fix threshold for locus call at 50 x which suitably worked even when low amounts of degraded DNA (12 pg) were investigated. In these analytical conditions, in fact, 25 PCR cycles allowed the genotyping of about 50 % and 35 % of the autosomal and the Y-specific markers on average, respectively, for each single amplification with a negligible frequency of drop ins (0.01 %). On the other hand, drop out artefacts reached 18-23 % when low copy number and degraded DNA samples were studied, with surviving alleles showing more than 600 reads in 2.9 % of the cases. Our data pointed out that the Precision ID Identity Panel allowed accurate typing of almost any amount of good quality/moderately degraded DNA samples, in duplicate tests. The analysis of low copy number DNAs evidenced that the same allele of a heterozygous genotype could be lost twice, thus suggesting that a third amplification could be useful for a correct genotype assignment in these peculiar cases. Using the consensus approach, a limited number of genotyping errors were computed and about 37 % of the autosomal markers was finally typed with a corresponding combined random match probability of at least 1.6 × 10-13, which can be considered an excellent result for this kind of challenging samples. In the end, the results presented in this study emphasize the crucial role of the expert opinion in the correct evaluation of artefacts arising from PCR-MPS technology that could potentially lead to genetic mistyping.

Deep knowledge of the genetic features of SARS-CoV-2 is essential to track the ongoing pandemic through different geographical areas and to design and develop early diagnostic procedures, therapeutic strategies, public health interventions, and vaccines. We describe protocols and first results of the Ion AmpliSeq™ SARS-CoV-2 Research Panel by a massively parallel sequencing (MPS) assay. The panel allows for targeted sequencing by overlapping amplicons, thereby providing specific, accurate, and high throughput analysis. A modified reverse transcription reaction, which consists of the use of a SARS-CoV-2 specific primers pool from the Ion AmpliSeq SARS-CoV-2 Research Panel, was assessed in order to promote viral RNA specific reverse transcription. The aim of this study was to evaluate the effectiveness of the Ion AmpliSeq™ SARS-CoV-2 Research Panel in sequencing the entire viral genome in different samples. SARS-CoV-2 sequence data were obtained from ten viral isolates and one nasopharyngeal swab from different patients. The ten isolate samples amplified with 12 PCR cycles displayed high mean depth values compared to those of the two isolates amplified with 20 PCR cycles. High mean depth values were also obtained for the nasopharyngeal swab processed by use of a target-specific reverse transcription. The relative depth of coverage (rDoC) analysis showed that when 12 PCR cycles were used, all target regions were amplified with high sequencing coverage, while in libraries amplified at 20 cycles, a poor uniformity of amplification, with absent or low coverage of many target regions, was observed. Our results show that the Ion AmpliSeq SARS-CoV-2 Research Panel can achieve rapid and high throughput SARS-CoV-2 whole genome sequencing from 10 ng of DNA-free viral RNA from isolates and from 1 ng of DNA-free viral RNA from a nasopharyngeal swab using 12 PCR cycles for library amplification. The modified RT-PCR protocol yielded superior results on the nasopharyngeal swab compared to the reverse transcription reaction set up according to the manufacturer’s instructions.

A set of eighty-two forensic samples with different levels of degradation, as well five in vitro damaged samples were analyzed by the Precision ID Identity Panel. PCR amplifications were performed with scalar amount of DNA (from 1 ng to 12 pg) and through different number of cycles. A minimum coverage of 50 x was adopted for "locus call". Very informative profiles (based on about 65-70% of the loci) were obtained even in highly degraded samples when the amount of template range from 0.1 to 1.0 ng. When dealing with low amount of degraded DNAs, no more than half of the loci were typed, and the risk of mistyping (due to drop out phenomena) increased dramatically. The employment of a high number of PCR cycles is discussed.

Osteogenesis imperfecta (OI) is a rare disease of collagen synthesis causing bone fragility. Also called “glass bone disease” since it manifests as spontaneous fractures, it is classified into nine types, both with dominant and recessive transmission. In 95% of cases OI is caused by mutations in COL1A1 and COL1A2 genes encoding the alpha1 and alpha2 chains of type 1 collagen, mainly null variants caused by frame-shift/nonsense mutations or splicing defects. In infants the differential diagnosis include not-accidental trauma, so child abuse. Families suspected of abuse often provide an unverified history of frequent fractures; conversely, the family history of individuals with OI often does not reveal any other affected individuals because of a de novo pathogenic variant in the proband or the presence of a mild phenotype in relatives. Therefore, legal medicine unit with DNA lab is crucial in these cases since it could early collect living or autopsy samples when a child abuse is suspected and then test DNA. We set up a MPS (massively parallel sequencing) panel including the coding regions of COL1A1 and COL1A2 and other 11 genes known to cause OI. We presented a case of suspected abuses in 2-month-old baby. MPS libraries were sequenced by Ion Torrent PGM platform; pathogenic variants and VUS (variants of uncertain significance) were confirmed by Sanger sequencing and familial segregation study was performed to better characterize the clinical significance of the mutation. This study remarks that MPS could help not only for identification, ancestry/phenotyping or molecular autopsy applications but also for forensic investigation over child abuse. The usefulness of this assay for diagnostic projects on victims of abuse together with post-mortem cases is discussed.

The HIrisPlex-S system, targeting a total of 41 SNPs, allows the simultaneous eye, hair and skin color prediction from DNA. In the present study, we developed a massive parallel sequencing (MPS) multiplex assay in order to genotype all the HIrisPlex-S markers in degraded casework samples. PCR amplicons sizes of target regions were kept below 180 bp, in order to allow analysis of degraded DNA samples. Individuals with known phenotype, artificially degraded DNA samples and a set of 2800M control DNA dilutions were sequenced on a Ion PGM System, in order to evaluate the concordance testing results and the forensic suitability of this 41-plex MPS assay. Full and reliable profiles could be obtained with 0.1 ng of input DNA. The increment of the number of PCR cycles results in improvement of sensitivity or in typing results but an increase of artifacts were also observed.

An inconsistency in the nomenclature used for the rapidly mutating (RM) Y-chromosomal short tandem repeat (Y-STR) marker DYS449 was noted in the above paper [[1]Robino C. Ralf A. Pasino S. De Marchi M.R. Ballantyne K. et al.Development of an Italian RM Y-STR haplotype database: results of the 2013 GEFI collaborative exercise.Forensic Sci. Int. Genet. 2015; 15: 56-63Abstract Full Text Full Text PDF PubMed Scopus (35) Google Scholar] In this paper [[1]Robino C. Ralf A. Pasino S. De Marchi M.R. Ballantyne K. et al.Development of an Italian RM Y-STR haplotype database: results of the 2013 GEFI collaborative exercise.Forensic Sci. Int. Genet. 2015; 15: 56-63Abstract Full Text Full Text PDF PubMed Scopus (35) Google Scholar], the DYS449 allele nomenclature introduced by Ballantyne et al. [[2]Ballantyne K.N. Keerl V. Wollstein A. Choi Y. Zuniga S.B. et al.A new future of forensic Y-chromosome analysis: rapidly mutating Y-STRs for differentiating male relatives and paternal lineages.Forensic Sci. Int. Genet. 2012; 6: 208-218Abstract Full Text Full Text PDF PubMed Scopus (196) Google Scholar] was used, instead of that described by Redd et al. [[3]Redd A.J. Agellon A.B. Kearney V.A. Contreras V.A. Karafet T. et al.Forensic value of 14 novel STRs on the human Y chromosome.Forensic Sci. Int. 2002; 4: 97-111Crossref Scopus (140) Google Scholar] and subsequently adopted by the International RM Y-STR User Group [[4]Ballantyne K.N. Ralf A. Aboukhalid R. Achakzai N.M. Anjos M.J. et al.Towards male individualization with rapidly mutating Y-chromosomal STRs.Hum. Mutat. 2014; 35: 1021-1032Crossref PubMed Scopus (133) Google Scholar] and in the AMPFlSTR® YFiler Plus kit [[5]Mulero J. Ballantyne J. Ballantyne K. Budowle B. Coble M. et al.Nomenclature update and allele repeat structure for the markers DYS518 and DYS449.Forensic Sci. Int. Genet. 2014; 13: e3Abstract Full Text Full Text PDF PubMed Scopus (4) Google Scholar]. To convert from the first [1Robino C. Ralf A. Pasino S. De Marchi M.R. Ballantyne K. et al.Development of an Italian RM Y-STR haplotype database: results of the 2013 GEFI collaborative exercise.Forensic Sci. Int. Genet. 2015; 15: 56-63Abstract Full Text Full Text PDF PubMed Scopus (35) Google Scholar, 2Ballantyne K.N. Keerl V. Wollstein A. Choi Y. Zuniga S.B. et al.A new future of forensic Y-chromosome analysis: rapidly mutating Y-STRs for differentiating male relatives and paternal lineages.Forensic Sci. Int. Genet. 2012; 6: 208-218Abstract Full Text Full Text PDF PubMed Scopus (196) Google Scholar] to the second [3Redd A.J. Agellon A.B. Kearney V.A. Contreras V.A. Karafet T. et al.Forensic value of 14 novel STRs on the human Y chromosome.Forensic Sci. Int. 2002; 4: 97-111Crossref Scopus (140) Google Scholar, 4Ballantyne K.N. Ralf A. Aboukhalid R. Achakzai N.M. Anjos M.J. et al.Towards male individualization with rapidly mutating Y-chromosomal STRs.Hum. Mutat. 2014; 35: 1021-1032Crossref PubMed Scopus (133) Google Scholar, 5Mulero J. Ballantyne J. Ballantyne K. Budowle B. Coble M. et al.Nomenclature update and allele repeat structure for the markers DYS518 and DYS449.Forensic Sci. Int. Genet. 2014; 13: e3Abstract Full Text Full Text PDF PubMed Scopus (4) Google Scholar] nomenclature, a simple correction factor needs to be applied, i.e. subtracting one from the repeat number of every DYS449 allele as described elsewhere [[5]Mulero J. Ballantyne J. Ballantyne K. Budowle B. Coble M. et al.Nomenclature update and allele repeat structure for the markers DYS518 and DYS449.Forensic Sci. Int. Genet. 2014; 13: e3Abstract Full Text Full Text PDF PubMed Scopus (4) Google Scholar]. The only consequence of this DYS449 allele nomenclature change is for comparing the RM Y-STR haplotypes newly obtained for the Italians in this study [[1]Robino C. Ralf A. Pasino S. De Marchi M.R. Ballantyne K. et al.Development of an Italian RM Y-STR haplotype database: results of the 2013 GEFI collaborative exercise.Forensic Sci. Int. Genet. 2015; 15: 56-63Abstract Full Text Full Text PDF PubMed Scopus (35) Google Scholar], with those from 111 worldwide populations obtained previously using a different nomenclature [[4]Ballantyne K.N. Ralf A. Aboukhalid R. Achakzai N.M. Anjos M.J. et al.Towards male individualization with rapidly mutating Y-chromosomal STRs.Hum. Mutat. 2014; 35: 1021-1032Crossref PubMed Scopus (133) Google Scholar]. Considering the DYS449 nomenclature correction, the complete absence of full 13 loci RM Y-STR haplotype matches in this dataset was confirmed. However, when limiting the haplotype comparisons to 11 RM Y-STR loci by excluding the multi-copy markers DYF399S1 and DYF403S1, the previously reported single haplotype match between a single Italian sample from Abruzzo and a single Austrian sample from Salzburg [[1]Robino C. Ralf A. Pasino S. De Marchi M.R. Ballantyne K. et al.Development of an Italian RM Y-STR haplotype database: results of the 2013 GEFI collaborative exercise.Forensic Sci. Int. Genet. 2015; 15: 56-63Abstract Full Text Full Text PDF PubMed Scopus (35) Google Scholar] disappeared when applying the corrected DYS499 nomenclature due to an observed mismatch at DYS449. Moreover, no additional haplotype match between Italians and worldwide populations was seen when considering these 11 RM Y-STR loci after DYS449 nomenclature harmonization. The corrected Supplementary Fig. S1 graphically depicting RM Y-STR haplotype matches in the tested populations is provided here. The corrected Supplementary Tables showing the results according to the current DYS449 nomenclature [3Redd A.J. Agellon A.B. Kearney V.A. Contreras V.A. Karafet T. et al.Forensic value of 14 novel STRs on the human Y chromosome.Forensic Sci. Int. 2002; 4: 97-111Crossref Scopus (140) Google Scholar, 4Ballantyne K.N. Ralf A. Aboukhalid R. Achakzai N.M. Anjos M.J. et al.Towards male individualization with rapidly mutating Y-chromosomal STRs.Hum. Mutat. 2014; 35: 1021-1032Crossref PubMed Scopus (133) Google Scholar, 5Mulero J. Ballantyne J. Ballantyne K. Budowle B. Coble M. et al.Nomenclature update and allele repeat structure for the markers DYS518 and DYS449.Forensic Sci. Int. Genet. 2014; 13: e3Abstract Full Text Full Text PDF PubMed Scopus (4) Google Scholar] for control DNA 2800M included in the PowerPlex® Y23 kit (Promega) and for the entire Italian dataset are provided here in Table S2 and S3, respectively. No other findings and conclusions were affected by the DYS449 nomenclature correction noted here. The following are Supplementary data to this article: Download .pdf (.07 MB) Help with pdf files Download .doc (.03 MB) Help with doc files Download .xlsx (.39 MB) Help with xlsx files Development of an Italian RM Y-STR haplotype database: Results of the 2013 GEFI collaborative exerciseForensic Science International: GeneticsVol. 15PreviewRecently introduced rapidly mutating Y-chromosomal short tandem repeat (RM Y-STR) loci, displaying a multiple-fold higher mutation rate relative to any other Y-STRs, including those conventionally used in forensic casework, have been demonstrated to improve the resolution of male lineage differentiation and to allow male relative separation usually impossible with standard Y-STRs. However, large and geographically-detailed frequency haplotype databases are required to estimate the statistical weight of RM Y-STR haplotype matches if observed in forensic casework. Full-Text PDF