Cell-type-dependent expression of a somatic tri-allelic pattern at D18S51 and Penta D: The statistical interpretation of forensic evidence in sexual assault investigations.

Laos, Khmer, Kui, and Yer ethnic populations had a high prevalence of α+-thalassemia, but the genetic background remains poorly understood. This study examined genetic variation at the α-globin gene cluster and three VNTR loci (D1S80, D17S5, and TPO) in these ethnic populations. For α-globin haplotype analysis, 110 subjects with normal α-globin and 232 subjects with α+-thalassemia were selected to analyze six polymorphic sites using the PCR-RFLP technique. For VNTR loci analysis, 447 subjects were examined for D1S80, D17S5, and TPO allele frequencies using a PCR-based method. The results of this study revealed that the most frequent haplotypes found in Laos, including (framework 1; + M - + − 0) linked to (-α3.7) and (framework 3; - S - + + -) related to (αCSα) were different from those found in Khmer and Kui [(framework 2; + S - + − 0) linked to (-α3.7) and (framework 1; + M - + + -) related to (αCSα)]. For the (αPSα) gene, the haplotype (+ S - + - -) of framework 2 was found in all ethnic groups, and the haplotype (- M - + + -) of framework 1 in the Yer only. The distribution of allele frequencies for the D1S80, D17S5, and TPO VNTR loci showed extensive genetic variation in the ethnic population studied. The number of alleles is higher than that of the previously reported populations. Based on D17S5 analysis, the phylogenetic tree suggested that Khmer and Kui ethnic groups had a close relationship but were distant from Laos. In addition, ethnic relationships were observed in the Yer and Kui populations. In contrast, consistent results were not obtained based on D1S80 and TPO analysis. The findings indicate genetic variation in these ethnic populations, but the conclusion remains tentative. However, this study provides useful information to better understand genetic origins and ethnic relationships in the region.

In recent years, DNA analysis techniques have drastically increased in sensitivity, allowing for low template DNA samples to be more easily detected and used for identification. Since the problems inherent in low template DNA are exacerbated in DNA mixture samples, it would be advantageous to incorporate an assay earlier in the DNA workflow that could detect a mixture and, potentially, determine the number of contributors. Some real-time PCR instruments have high-resolution melt curve analysis (HRM) capabilities, allowing for an opportunity to integrate an HRM screening assay into a commercial DNA quantification kit. This work describes the integration of a mixture screening HRM assay using STR loci D5S818 and D18S51 into Qiagen’s Investigator Quantiplex® kit. The integrated Quantiplex®-HRM assay was tested on two qPCR platforms: The Rotor-Gene® Q and the QuantStudio™ 6 Flex. Data from this assay was analyzed using linear discriminant and support vector machine analyses for sample classification. When HRM curve data from the Rotor-Gene® Q was used for classification, the integrated assay exhibited an overall accuracy of 89.39%, correctly classifying 87.5% of single source samples and 100% of mixtures. When HRM curve data from the QuantStudio™ 6 Flex was used for classification, the integrated assay exhibited an overall accuracy of 87.88%, correctly classifying 87.5% of single source samples and 90% of mixtures. The overall accuracy of the integrated Quantiplex®-HRM assay on both instruments met our goal of ≥ 80% accuracy, demonstrating the viability of the assay to detect mixtures when integrated into a commercial quantification kit.

During paternity investigation with Identifiler™ set of autosomal Short Tandem Repeats (STRs), a genetic mismatch was observed with D7S820 between the disputed father and child. The genotype at this locus in the disputed father, mother and child was 10/10, 11/12 and 11/11, respectively. The combined paternity index and probability of paternity after including the mutation in the calculation were 7.6×107 and 0.9998, respectively. Both values supported the suspicious father as the biological father of the child. Further analysis of Y-STRs revealed matching of all the alleles of the child with that of the suspicious father. It suggested that the mismatch at the D7S820 locus might be a case of mutation. DNA sequencing of D7S820 PCR products of the child and both the parents helped in determining that the child inherited the expanded repeat of the paternal allele 10, which was transmitted as allele 11 in the child from the suspicious father.

There is a little investigation of the genetic diversity of the Libyan population. This study aimed to explore STR loci in the Libyan western mountain population to analyse its genetic landscape. Allele frequency for 15 autosomal STR loci (D8S1179, D21S11, D7S820, CSF1PO, D3S1358, THO1, D13S317, D16S539, D2S1338, D19S433, VWA, TPOX, D18S51, D5S818, FGA) included in the Identifiler®Plus kit were investigate in 120 random unrelated individuals from the Westerner Mountain area of Libya. No deviations from Hardy-Weinberg equilibrium were seen, with the exception of D8S1179, D21S11, D3S1358, TH01 and FGA loci. A total of 129 alleles were observed with the corresponding allelic frequencies ranging between 0.00417 and 0.47917 of the studied sites were observed, and that alleles 8 and 12 are the highest frequency for all studied areas for the TPOX and D5S818 sites respectively had a value of (0.47917). D18S51 was also observed to be the most diverse site (Expected heterozygosity: 0.88312 and probability of matching: 0.0354). The results indicate that the 15 STR loci have a significant genetic diversity among studied individuals and are suitable for personal identification in the criminal field, and the importance of this work lies in the creation of a database of frequency distribution of allelic frequencies to be used for criminal identification, identification of missing persons and paternity tests.

Background/Objectives: Short tandem repeat (STR) loci are widely used in forensic genetics for identification and kinship analysis. Traditionally, these loci were selected to avoid medical associations, but recent studies suggest that loci such as TH01 and D16S539 may be linked to psychiatric conditions like schizophrenia. This study explores these potential associations and considers the privacy implications related to disease susceptibility. Methods: We analyzed 19 STR loci, including CODIS core loci and additional loci like Penta D and Penta E. Statistical analyses were conducted on a dataset of schizophrenia patients and matched control individuals to assess the relationship between STR polymorphisms and schizophrenia risk. Results: No significant associations were found between the 19 analyzed loci and schizophrenia in this dataset. While initial analyses revealed minor allele frequency differences at the D3S1358, D13S317, and TPOX loci between the schizophrenia and control groups, these differences did not retain statistical significance following Bonferroni correction (corrected p < 0.0026 for all loci). Conclusions: Although no significant associations were found between STR loci and schizophrenia, this study highlights the importance of considering the potential for forensic DNA data to reveal health-related information. As forensic DNA databases continue to expand, there is a growing need to reassess ethical and legal guidelines to ensure the protection of individual privacy. Future research should continue exploring these genetic associations with larger, more diverse samples to further understand their implications.

الفعل الاجرامي هو سلوك غير قانوني وقد يكون جريمة جنائية ، ويستخدم المجرمون طرقا مختلفة لاخفاء الادلة في مسرح الجريمة لكن غالبا ما يجد خبراء الطب الشرعي اثار الحمض النووي على العناصر الموجودة في مسرح الجريمة, ومن هذه الادلة هو الجيتار. والجيتار من الالات الموسيقية التي يعزف عليها البشروالتي يمكن استخدامها للمقارنة . استخدمت الطرق في هذه الدراسة 18 عينة من أوتار الجيتار التي تم استخدامها لمدة خمس دقائق وحضنت في درجة حرارة الغرفة. تم بعد ذلك تقسيم العينات الـ 18 إلى ثلاث مجموعات، حيث تتكون كل مجموعة من ست عينات وتم تحضينها لمدة يوم وخمسة و10 أيام. تم بعد ذلك قياس الحمض النووي باستخدام مطياف الاشعة فوق البنفسجية للحمض النووي المستخلص بطريقة DNAzol . و كان معدل تقدير الحمض النووي في بعد الحضن لليوم الاول ( 152.57 ± 48.02 نانوغرام/ميكروليتر) واليوم 5 كانت ( 138.66 ± 47.66 نانوغرام/ميكروليتر)، واليوم 10 كانت ( كانت 87.09 ± 9.07 نانوغرام/ميكروليتر) . وتم الكشف عن كفاءة الحمض النووي المستخلص للمقارنة بتضخيم 3 مواقع هي D18S51 و FES و D8S1179 بتنقية تفاعل البلمرة المتسلسل، حيث ظهرت الحزم ذوات الاحجام ( 290-366) و((222 – 250 و( 203 – 224) زوج قاعدي, على التوالي. ونستنتج امكانية استخدام اوتار الجيتار كمصدر لعزل الحمض النووي بعد وقوع الجريمة بعدة ايام وبدرجة حرارة الغرفة.

Background: Generally, the primary sample used for DNA testing comes from blood.Improvements in existing analytical methodologies can reduce the need for DNA levels used in examinations so that samples used can come from other parts of the body with retrieval methods that are non-invasive, more straightforward, easier to store, and more resistant to decay, for example, nails.Materials and methods: Nail clipping samples were taken from the hands and feet of an unidentified corpse, which were then divided into 5 groups: the control group without treatment, the soaking group with fresh water and salt water on days 3 and 7. DNA extraction using chloroform-phenol, measurement of DNA levels, and purity using NanoDrop spectrophotometer.Each sample representative with the lowest and highest levels was further analyzed at D18S51 loci.Results: Average levels (g/dl SD) of DNA nail clippings in the control group 299.68 44.58, fresh water immersion group day 3 of 555.14 192.12 and 7 of 429.82 66.65, salt water immersion group day 3 of 228.04 81.89 and day 7 of 400.76 53.30.Polymerase Chain Reaction (PCR) visualization results from D18S51 loci show DNA bands or bands appear all 100%, with PCR product sizes of 290-366 bp.Conclusion: Quantitatively, there was no effect of decreasing DNA levels and purity on fresh and saltwater immersion, but qualitatively, DNA bands appeared thinner in the immersion group in fresh water.

Introduction: Social interactions such as marriage patterns in communities descended from the Tengger ethnic group are hypothesized to affect genetic variations that occur in communities descended from the Tengger ethnic group in the Tengger highland area. DNA identification using Short Tandem Repeat (STR) is a reference for assessing genetic variations that exist in populations. This research aims to determine allele frequencies and genetic variations in the Tengger tribe in East Java, Indonesia. Method: This observational study used 50 from the Tengger tribe. DNA isolation was done using the Promega kit, PCR was done using 23 autosomal STR loci, and visualization was done using Capillary electrophoresis to obtain allele data from all loci. The data is analyzed and then presented using tables and graphs. Result: The results show various allele frequencies and the D10S1248 locus on allele 13 has the highest frequency with 0.5300. The percentage of homozygous and heterozygous alleles shows that the CSF1PO locus has a homozygous allele percentage of 42% and the D21S11 and SE33 loci have a heterozygous allele percentage of 94%. The highest Heterozygosity value is found at the SE33 locus with a value of 0.9098 and the highest Homozygosity value is at the CSF1PO locus with a value of 0.3864. Conclusion: The various allele frequencies and forensic data obtained conclude that the Tengger tribe has unique characteristics.

BACKGROUND Adipose tissue is often overlooked in DNA testing due to misconceptions about its DNA content. However, its shock-absorbing qualities may be useful for high-pressure scenarios like bomb blasts. This study aimed to evaluate DNA quality and quantity in adipose tissue affected by blasts compared to that in unaffected tissue.&#x0D; METHODS 10 adipose tissue samples were taken from regions near and far from the blast, representing the blast-exposed and non-blast-exposed groups. The adipose tissue was stored at a low temperature for 5 days, after which an organic extraction method was applied. The purity of the DNA extract was assessed using a NanoDrop spectrophotometer, and its integrity was evaluated using 0.8% concentration gel electrophoresis at 60 V for 90 min. DNA typing was conducted using the GlobalFiler™ kit, and DNA quantity was determined with the Quantifiler™ Trio DNA Quantification kit.&#x0D; RESULTS Of 20 DNA extracts from adipose tissue, all samples demonstrated purity, integrity, and complete typing results. Adequate integrity was found in 90% of samples in both groups. A 50% incidence of allele shifting was observed at the D7S820 locus within the blast-exposed group.&#x0D; CONCLUSIONS DNA from blast-exposed adipose tissue exhibited no significant quality or quantity differences from non-blast-exposed tissue. This suggested adipose tissue’s potential as an alternative DNA source in a bomb explosion.

Short tandem repeat (STR) typing has been regularly used in paternity disputes and forensic human identification linked caseworks. Occasionally, forensic scientists come across aberrant allele patterns during STR typing because of mutations, genetic variations, and other abnormalities. The tri-allelic pattern of STR is rare, particularly, the case where this pattern exists at 4 loci. Here, we report the type II tri-allelic patterns observed at vWA, SE33, D8S1179, and D13S317 loci in the product of conception (POC) sample during the course of our regular paternity case investigation. The DNA extracted from the blood samples and tissue of POC were subjected to STR typing for autosomal and sex STR loci using the commercial QIAGEN's Investigator® IDplex Plus Kit and QIAGEN's Investigator® 24plex QS Kit. Capillary electrophoresis was carried out in 3500 and 3500xL Genetic Analyzer Applied Biosystems and genotyped using GeneMapper ID-X Software v1.5 and v1.6. In this case of paternity inclusion, the POC sample displayed type II tri-allelic patterns at vWA (16, 19, 20), SE33 (19, 28.2, 29.2), D13S317 (16, 19, 20), and D8S1179 (10, 13, 17) loci. In addition, the POC displayed an abnormal genotype with a heterozygous peak imbalance (type II-B) of (1:2) pattern at D3S1358, D21S11, and D16S539 loci, of (2:1) pattern at D1S1656, D12S391, D10S1248, D2S1338, D2S441, D18S317, FGA, CSF1PO, and D5S818 loci, and type II-C allelic pattern (one single peak with triplicate height) at D19S433 and DS7820 loci. Understanding of such anomalous genotypes improves the knowledge about tri-allelic pattern of CODIS loci and helps in the appropriate interpretation of the results in STR typing.

DNA typing based on short tandem repeat (STR) analysis is an effective forensic method for human identification. Some STRs are contained within the introns of protein-coding genes and are transcribed as pre-mRNAs. However, the possibility of using RNA for STR analysis is yet to be fully explored. Considering that RNA in forensic samples is relatively stable, especially under dry- and low-temperature conditions, we hypothesized that STR information could be obtained from RNA. Here, we investigated the possibility of conducting RNA-based STR analysis using the D18S51 locus as a model.

The DNA within the Human Genome involves loci with high polymorphism such as short tandem repeats (STR). The STR polymorphism analysis is extensively used for numerous purposes such as identification studies in forensic sciences, paternity tests, genome mapping, and population studies. In this study, blood and swab samples were obtained from 200 unrelated healthy individuals living in Elazig province. DNA was amplified via the AmpFlSTR Identifiler PCR Amplification Kit (Applied Biosystems, USA) to analyze the 15 STR loci. Using the ABI PRISM310 Genetic Analyzer capillary electrophoresis, the samples were analyzed and the allelic ladder was used to compare and identify the alleles of the samples, the distribution of the allele frequencies and peak values were examined. For statistical analysis Arlequin v.3.11 and PowerStats v.1.2 were used to assess the data. To determine if the allele frequencies were compatible with the Hardy-Weinberg (HW) equilibrium, the Markov chain and the Fisher's exact test p-value (p&lt;0.05) were examined. It was determined that the D8S1179 (0.00002), D21S11 (0.00593) and vWA (0.04939) loci were not compatible with the HW equilibrium, but the rest of the loci were compatible. For the loci, the combined power of exclusion is 0.9999995, the combined power of discrimination is 0.9999999999998. The probability of matching for the 15 STR loci was calculated to be 1 in 1.364x10ˉ17. The power of discrimination was quite high when all the loci were considered. The results of this study indicate that the populations residing in the Elaziğ province may benefit from paternity testing, forensic identification, and genomic mapping using the data as an appropriate marker.

DNA identification has an important role in forensics to investigate a case. However, the problem that is often encountered is that the DNA found is difficult to analyze because the amount is relatively small and degraded. An alternative used in DNA examination is to design mini primers for amplification. Mini primers can produce DNA profiles even in samples that have very low quantity and quality. Therefore, this study aims to determine the effectiveness of mini primers for amplification of TH01 and D21S11 loci in DNA exposed to high temperatures. In addition, to determine the effect of high temperature exposure on DNA quality and quantity. The samples used were 3 ml of human blood treated with 100, 150, and 200ºC for 10, 20, and 30 minutes. The method used was PCR amplification using mini primers TH01 and D21S11 loci. Quantity data were analyzed by a parametric test using the Two Way Anova method. Meanwhile, DNA quality was tested by agarose gel electrophoresis. The results showed that temperature treatment affected the quantity and quality of DNA. The TH01 locus in DNA samples treated with high temperature can be amplified using primer pairs MP1FTH01 and MP1RTH01. At the D21S11 locus, the use of primers 1 and 2 can be combined to identify DNA samples with high temperature treatment

Short tandem repeats (STRs) are repeating DNA sequences used in forensic human identity testing and the diagnosis of aneuploidies. Many STRs like Penta D and TPOX are used routinely for paternity tests, but these tests are not widely used in sub-Saharan Africa. In this study we recruited individuals from Gabonese families seeking a paternity test. After DNA extraction from buccal swabs, we genotyped samples using a panel of 22 STRs. A total of 115 unrelated subjects from 39 families were included. Allele frequencies of the 22 STR loci were determined in unrelated Gabonese subjects. The most polymorphic loci were D21S11 (16 alleles) and FGA (17 alleles), while D3S1358 and TH01 loci were less polymorphic, with five alleles each. Deviation from Hardy–Weinberg equilibrium was observed for TPOX, D3S1358, CSFPO and D7S820 loci. We reported tri-allelic patterns that indicate aneuploidies at a combined frequency of 4% (4/115) with 3% for Penta D (1/35) and 3% for TPOX (3/102). Furthermore, we identified a new tri-allelic genotype 5-8-16 for the Penta D locus located on chromosome 21 in a healthy subject. In addition, we observed three tri-allelic variants of TPOX, located on chromosome 2, in healthy subjects, namely 8-10-11, 8-9-10, and 8-8-10. Our study revealed unsuspected polymorphic variations in Penta D and TPOX for the first time in Gabon, raising several questions about chromosomal disorders. Further population genetics studies are needed in Gabon to better characterize these variations, both qualitatively and quantitative.

Abstract Background: The role of DNA analysis for ethnicity inferencing is a topic that attracts much interest from researchers in forensic identification, especially for identifying unknown bodies and trace evidence. So far, the approaches considered effective for ethnic inferencing are autosomal single-nucleotide polymorphisms, Y-chromosome short-tandem repeats (STRs), and mitochondrial DNA haplotyping, which successfully demonstrates the association of specific nucleotides or patterns with population groups. Ethnic inferencing based on autosomal STRs is complex due to the nature of recombination in gamete formation. Aim: This study attempts to use clustering analysis to associate alleles and loci of autosomal STRs with population groups. Materials and Methods: We examined the allele frequency data from 19 STRs loci from the Malay Indonesian population (n = 470) to compare with other populations, namely, Chinese Indonesian (n = 133) and four reference populations (Malay Malaysian, Filipino, Chinese, and Caucasian). K-Medoids clustering analysis was carried out to pinpoint alleles and loci affecting the population clustering process. Results: The first stage of clustering results placed Malay Indonesians and four other Asian populations, namely, Chinese Indonesian, Malay Malaysian, Filipino, and Chinese, in Cluster 1, whereas the Caucasian group was in Cluster 2. It indicates that the CSF1PO, D5S818, and D8S1179 loci significantly distinguished the five Asian population groups from the Caucasian group, whereas D2S441, D8S1179, and D22S1045 were the three loci that significantly influenced the separation between Malay Indonesians and other groups. Conclusions: We conclude that K-medoids clustering analysis has the potential to play a role in ethnicity estimation by pinpointing specific STRs alleles.

INTRODUCTION: We focused on a sample size of 141 unrelated Rwandan persons to genotype21 STR loci that were relied up in establishing allele frequencies, heterozygosity and power ofexclusion. This study aims at exploring allele frequencies on a representative sample from Rwandanpopulation to determine probability of paternity for sampled families basing on polymorphic STRsloci, using 21 autosomal-STR loci by Genetic Analyzer 3500X.METHODS: This was an experimental study and global filer TM Express PCR Amplification kit wasused to amplify 21 autosomal STR loci.RESULTS: The total number of observed alleles was 270; the largest number of different alleleswas seen in SE33 and D18S51 loci. The locus with the highest heterozygosity was SE33, while locusTH01 had the lowest heterozygosity. The heterozygosity of the 21 STR loci ranged from 71.3%(TH01) to 91.6% (SE33) with an average of 81.1% a good indicator of high genetic variability. Forall microsatellites analyzed the power of exclusion ranged from 43.4% (TH01) to 78.1% (SE33)with an average of 58.2%. For seven of eight cases examined in paternity test alleged father wasnot excluded as biological father of child. The results found in examination of case 8 indicated thatthe alleged father was not the biological father of the child.CONCLUSION: Based on calculated statistical parameters, the population of Rwanda may usethese 21 STR loci as a vital tool for forensic identification and paternity testing.

Capillary electrophoresis is widely used to study short tandem repeats (STRs) in forensic genetics. However, next-generation sequencing platforms have become a new strategy for forensic DNA typing. In this study, we report a false four-step STR mutation between an alleged father (AF) and child in a paternity case. A total of 23 autosomal STR loci were evaluated using the Huaxia™ Platinum and Goldeneye™ 20A kits, revealing a single mismatch in D8S1179 between the AF (10/10) and the male child (14/14). Additional Y-STR typing of the AF and child was performed, and the results were consistent with those based on 27 Y-STR loci. To further confirm the experimental results, we sequenced the individuals using the MiSeq FGx system and detected 10/15 unbalanced alleles in the D8S1179 locus of the AF and 14/15 unbalanced alleles in the D8S1179 locus of the child. Sanger sequencing revealed that both the AF and child had the C→G point mutation in the primer binding region of D8S1179 resulting in allelic dropout. Therefore, the verification of STR typing by different sequencing systems is helpful for the interpretation of results in cases of multistep STR mutations.

DNA can be obtained from all parts of the body with the same profile in each person. The oral mucosal epithelium is one of the DNA sources that is often used for individual identification because of taking it using a swab method that does not hurt the volunteers. The purpose of this study was to determine DNA polymorphism in Biology students of Class of 2022. The sample used in this study were 49 people with details, 34 female and 15 male students. After cheek mucosa was sampling, the PCR and electrophoresis was performed to see the DNA bands formed. The visible band was measured in length and calculated using a formula to see the repeatability of the D1S80 locus on the DNA band. From the 49 samples which were collected, the DNA bands were seen in 27 samples with DNA band lengths ranging from 400-600 bp. The number of bands in each sample were also different. The highest frequency of locus repetitions was 26 repetitions and the lowest was 14 repetitions.

Short tandem repeat (STR) automatic typing technology is extensively used in forensic laboratories with commercial kits, in rare cases genotyping misinterpretations or mislabeling may occur due to unexpected rare alleles. This study refers to the investigation of several rare alleles observed from routine cases. Besides cross-kit verification with Goldeneye 25A (Beijing PeopleSpot Inc, China) and Huaxia platinum (Thermo Fisher Scientific, USA) kits, the next-generation sequencing technology by MiSeq FGx System (Illumina, USA) was applied to further validation. To solve the inconsistent outcomes reached by the above mentioned approaches at D2S441 locus, single gene amplification, gene cloning, and genetic sequencing was also performed. As a result, five rare alleles were detected. Two novel alleles of allele 3 at the D13S317 locus and allele 5 at the D2S441 locus were found; three previously reported alleles of allele 9 at D1S1656 locus, allele 19 at Penta D locus, and allele 28 at D12S391 locus in STRBase were initially supplemented with sequence information. We, therefore, propose that such uncommon observations with rare events should be carefully investigated and interpreted.