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1.
Leg Med (Tokyo) ; 69: 102447, 2024 Apr 16.
Article in English | MEDLINE | ID: mdl-38640874

ABSTRACT

This study aimed to estimate A-STR mutation rates in 2,317 Korean parent-child trios by examining 20 Combined DNA Index System (CODIS) core loci (D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51, D21S11, CSF1PO, FGA, TH01, TPOX, vWA, D1S1656, D2S441, D2S1338, D10S1248, D12S391, D19S433, and D22S1045) and three non-CODIS loci (Penta E, Penta D, and SE33). Locus-specific mutation rate estimates varied from 0.00 to 8.63 × 10-3 per generation, with an average mutation rate of 1.62 × 10-3 (95 % CI, 1.39-1.88 × 10-3). We also combined data from previous studies to obtain comprehensive genetic values for the Korean population, and the average mutation rate was 1.59 × 10-3 (95 % CI, 1.38-1.82 × 10-3). Single-step mutations (95.69 %) and double-step mutations (3.35 %) were observed in the mutation pattern analysis, and cases expected to have multi-step mutations (0.96 %) were also observed. Large-sized alleles exhibited more loss mutations than gain mutations, and paternal mutations (62.68 %) were more frequently observed than maternal mutations (19.62 %). The calculated values and features of the 23 A-STRs explored in this study are expected to play a crucial role in establishing criteria for forensic genetic interpretation.

2.
J Forensic Sci ; 2024 Jan 26.
Article in English | MEDLINE | ID: mdl-38275209

ABSTRACT

The DNA intelligence tool, DNA methylation-based age prediction, can help identify disaster victims and suspects in criminal investigations. In this study, we developed a costal cartilage-based age prediction tool that uses massive parallel sequencing (MPS) of age-associated DNA methylation markers. Costal cartilage samples were obtained from 85 deceased Koreans, aged between 26 and 89 years. An MPS library was prepared using two rounds of multiplex polymerase chain reaction of nine genes (TMEM51, MIR29B2CHG, EDARADD, FHL2, TRIM59, ELOVL2, KLF14, ASPA, and PDE4C). The DNA methylation status of 45 CpG sites was determined and used to train an age prediction model via stepwise regression analysis. Nine CpGs in MIR29B2CHG, FHL2, TRIM59, ELOVL2, KLF14, and ASPA were selected for regression model construction. A leave-one-out cross-validation analysis revealed the high performance of the age prediction model, with a mean absolute error (MAE) and root mean square error of 4.97 and 6.43 years, respectively. Additionally, our model showed good performance with a MAE of 6.06 years in the analysis of data of 181 costal cartilage samples collected from Europeans. Our model effectively estimates the age of deceased individuals using costal cartilage samples; therefore, it can be a valuable forensic tool for disaster victim and missing person investigation.

3.
Genes (Basel) ; 14(11)2023 Oct 24.
Article in English | MEDLINE | ID: mdl-38002929

ABSTRACT

The study of gender markers is essential in forensic genetic analysis. Mutations in the X or Y homologs of the amelogenin gene can be misleading, resulting in serious mistakes in forensic genetic analysis. We recently discovered two male cases of the X homolog of the amelogenin (AMELX) allelic dropout while analyzing short tandem repeat genotypes obtained from crime scene evidence. Subsequently, we evaluated the molecular characteristics of AMELX allelic dropout in this study. We used two previously reported amelogenin primers to verify a half level of amelogenin gene amplification intensity in the two male cases, which we confirmed was caused by AMELX allelic dropout. We then characterized the point mutation using Sanger sequencing and designed mutation-specific primers that could overcome AMELX allelic dropout. Short tandem repeat genotyping analysis confirmed that the AMELX allelic dropout was recovered by the mutation-specific primer designed specifically for this case. The sequencing of the AMELX allele revealed a single-point variant from A→G at base position 7 downstream from the 3' end in the amelogenin forward primer-binding region. This point mutation was identically found in two different male cases, resulting in AMELX allelic dropout. To our knowledge, these mutations and the X homolog amplification failure of amelogenin have not been reported in the Korean population. Our study provides a reliable approach to AMELX allelic dropout due to rare case mutations and could enable the better interpretation of gender markers for forensic samples.


Subject(s)
Amelogenin , Point Mutation , Humans , Male , Alleles , Amelogenin/genetics , Asian People
4.
Forensic Sci Int Genet ; 65: 102875, 2023 07.
Article in English | MEDLINE | ID: mdl-37084624

ABSTRACT

Y-chromosomal short tandem repeats (Y-STRs) have been widely used in forensic genetics, and accurate knowledge of mutation rates at Y-STR loci is essential in kinship analysis. The main aim of this study was to estimate Y-STR mutation rates in Korean males. To obtain locus-specific mutations and haplotypes at 23 Y-STRs, we analyzed samples from 620 Korean father-son pairs. In addition, we also analyzed 476 unrelated individuals using the PowerPlex® Y23 System, with the aim of augmenting the available data for the Korean population. The PowerPlex® Y23 system facilitates analysis of the 23 Y-STR loci (DYS576, DYS570, DYS458, DYS635, DYS389 II, DYS549, DYS385, DYS481, DYS439, DYS456, DYS389 I, DYS19, DYS393, DYS391, DYS533, DYS437, DYS390, Y GATA H4, DYS448, DYS438, DYS392, and DYS643). Locus-specific mutation rate estimates varied from 0.00 to 8.06 × 10-3 per generation, with an average mutation rate of 2.17 × 10-3 (95% CI, 1.5-3.1 × 10-3). To obtain comprehensive genetic values for the Korean population, we combined data obtained in this study with previously reported values, thereby enabling us to estimate the locus-specific mutation rates regarding 22,711 allele transmissions. By combining these data, we obtained an overall average mutation rate of 2.91 × 10-3 (95% CI, 2.3-3.7 × 10-3). In addition, among the 476 unrelated Korean males, we detected 467 different haplotypes, with an overall haplotype diversity value of 0.9999. By extracting haplotypes of Y-STRs described in previous literature on 23 Y-STR reported in Korea, we obtained gene diversities for 1133 Korean individuals. We believe that the values and characteristics of the 23 Y-STRs analyzed in this study will contribute to establishing criteria for forensic genetic interpretation, including kinship analysis.


Subject(s)
Mutation Rate , Nuclear Family , Male , Humans , Haplotypes , Chromosomes, Human, Y , Microsatellite Repeats , Fathers , Republic of Korea , Genetics, Population , DNA Fingerprinting
5.
Article in English | MEDLINE | ID: mdl-36361134

ABSTRACT

The forward head posture of visual display terminal (VDT) users induces various physical and cognitive clinical symptoms. However, few studies have been conducted to identify and solve problems associated with VDT posture. This study aimed to examine the adverse effects of VDT posture and the positive effects of traction-combined workstations by measuring postural alignment, muscle properties, blood velocity, preference, and working memory. Thirty-four healthy VDT users (18 males and 16 females aged 20-30 years) participated in the experiment at three workstations, including conventional (VDT_C), head support (VDT_S), and upright (VDT_U) workstations. They conducted 2-back working memory task. The craniovertebral angle (CVA), muscle tone and stiffness, blood velocity and visual analogue discomfort scale (VADS) were measured to examine the influence of workstations. VDT_C showed increased muscle tone or stiffness in the levator scapulae (LS), suboccipital muscle (SM), and sternocleidomastoid muscle (SCM) and an increased reaction time (RT) in working memory. However, VDT_S showed decreased stiffness and tone of SM and improved comfort. In addition, VDT_U showed decreased stiffness or tone of the LS and SCM and improved blood velocity and RT. In conclusion, maintaining neutral alignment significantly improved working memory performance, muscle properties, and blood velocity.


Subject(s)
Computer Terminals , Superficial Back Muscles , Male , Female , Humans , Memory, Short-Term , Traction , Hemodynamics , Cognition , Weight Loss
6.
Front Psychiatry ; 13: 925823, 2022.
Article in English | MEDLINE | ID: mdl-36147982

ABSTRACT

Background: In animal experiments, the habenula and septal nuclei are known as the key brain areas of depression. However, there are few magnetic resonance imaging (MRI) studies on the functional connectivity between these areas and the subcortical areas in humans with major depression. We aimed to investigate the difference in resting-state functional connectivity (RSFC) among the major regions of interest (ROI) in the subcortical areas, including both the habenula and septal nuclei. Methods: We performed the seed-to-voxel analysis to investigate the RSFC between both the habenula and septal nucleus, as well as other subcortical regions. Furthermore, ROI-to-ROI analysis was performed among the combinations of ROI pairs in the subcortical areas. Results: The seed-to-voxel analysis showed a lower RSFC between the left habenula and the cerebellum in major depressive disorder (MDD) than in healthy controls (HCs). As a result of ROI-to-ROI analysis in subcortical areas, a total of 31 pairs of FCs in the MDD group showed a lower RSFC than in the HCs group. Conclusion: This study revealed a lower RSFC between the left habenula and cerebellum in patients with MDD and reduced RSFC among numerous subcortical areas. These new findings on the neural circuitry of MDD might contribute to an in-depth understanding of depression.

7.
Sensors (Basel) ; 22(16)2022 Aug 18.
Article in English | MEDLINE | ID: mdl-36015953

ABSTRACT

The diagnosis of small vessel disease is attracting interest; however, it remains difficult to visualize the microvasculature using 3 Tesla (T) magnetic resonance imaging (MRI). Therefore, this study aimed to visualize the microvascular structure and measure a slow flow on 3T MRI. We developed a microcirculation system using piezoelectric pumps connected to small tubes (0.4, 0.5, 0.8, and 1.0 mm) and evaluated various MR sequences and imaging parameters to identify the most appropriate acquisition parameters. We found that the system could image small structures with a diameter of 0.5 mm or more when using a 1 m-long tube (maximal signal intensity of 241 in 1 mm, 199 in 0.8 mm, and 133 in 0.5 mm). We also found that the highest signal-to-noise ratio (SNR) appeared on 2-dimensional time-of-flight low-resolution imaging and that the flow velocity (10.03 cm/s) was similar to the actual velocity (11.01 cm/s in a flowmeter) when velocity encoding of 30 cm/s was used in a 0.8 mm-diameter tube. In conclusion, this study demonstrates that a microcirculation system can be used to image small vessels. Therefore, our results could serve as a basis for research on vessels' anatomical structure and pathophysiological function in small vessel disease.


Subject(s)
Magnetic Resonance Imaging , Ultrasonics , Magnetic Resonance Imaging/methods , Microcirculation , Microvessels/diagnostic imaging , Phantoms, Imaging , Signal-To-Noise Ratio
8.
Sensors (Basel) ; 22(6)2022 Mar 16.
Article in English | MEDLINE | ID: mdl-35336474

ABSTRACT

This study aimed to develop a magnetic resonance imaging (MRI)-compatible flow delivery system and individualized models of circle of Willis (CoW), which include 50% and 100% blockage in internal carotid artery (ICA50 and ICA100), and 100% blockage in vertebral artery (VA100). Images were obtained using 3D time-of-flight and phase-contrast magnetic resonance angiography (MRA) sequences, and changes in velocity and flow direction at CoW models were analyzed. For the ICA50 and VA100 models, the flow was similar to that of the normal model. For the ICA 50 model, it was found that 50% blockage did not affect cerebral blood flow. For the VA100 model, decreased flow in the posterior cerebral artery and a change to the flow direction in the posterior communicating artery were found. For the ICA100 model, particularly, decreased flow in the ipsilateral middle and anterior cerebral arteries and a change to the flow direction in the ipsilateral anterior cerebral artery of the CoW were found. These results demonstrated that the flow system with various CoW disease models tailored to individual characteristics could be used to predict stroke onset more quickly. For the ICA50 and VA100 models, the possibility of cerebral infarction was significantly lower. On the other hand, for the ICA100 model, there was a high possibility of decreased flow, which could lead to cerebral infarction.


Subject(s)
Carotid Stenosis , Carotid Artery, Internal/diagnostic imaging , Cerebrovascular Circulation , Circle of Willis/diagnostic imaging , Circle of Willis/pathology , Humans , Magnetic Resonance Imaging
9.
Sensors (Basel) ; 21(19)2021 Sep 23.
Article in English | MEDLINE | ID: mdl-34640671

ABSTRACT

In this study, we proposed a novel pulse wave velocity (PWV) technique to determine cerebrovascular stiffness using a 3-tesla magnetic resonance imaging (MRI) to overcome the various shortcomings of existing PWV techniques for cerebral-artery PWV, such as long scan times and complicated procedures. The technique was developed by combining a simultaneous multi-slice (SMS) excitation pulse sequence with keyhole acquisition and reconstruction (SMS-K). The SMS-K technique for cerebral-artery PWV was evaluated using phantom and human experiments. In the results, common and internal carotid arteries (CCA and ICA) were acquired simultaneously in an image with a high temporal resolution-of 48 ms for one measurement. Vascular signals at 500 time points acquired within 30 s could generate pulse waveforms of CCA and ICA with 26 heartbeats, allowing for the detection of PWV changes over time. The results demonstrated that the SMS-K technique could provide more PWV information with a simple procedure within a short period of time. The procedural convenience and advantages of PWV measurements will make it more appropriate for clinical applications.


Subject(s)
Cardiovascular System , Pulse Wave Analysis , Brain/diagnostic imaging , Humans , Magnetic Resonance Imaging , Magnetic Resonance Spectroscopy
10.
J Forensic Sci ; 66(6): 2362-2368, 2021 Nov.
Article in English | MEDLINE | ID: mdl-34296771

ABSTRACT

Dogs (Canis lupus familiaris) are among the most common companion animals in the Republic of Korea. Recently, there have been many criminal cases of dog cruelty, injury, and theft, among others. This has increased the importance of dog-related biological evidence at crime scenes. The National Forensic Service of the Republic of Korea conducts short tandem repeat (STR) analysis using the Thermo Scientific Canine Genotypes™ Panel 2.1 Kit (Canine Kit) to identify individual dogs through forensic analysis. The Canine Kit was developed as a forensic STR kit for the identification of individual dogs. However, an allelic ladder was neither developed nor included in the commercial kit, leaving an issue of accurate genotyping. Primer details for the 18 markers used in the Canine Kit are proprietary information, and thus, unavailable to end-users. In this study, an allelic ladder was constructed with 160 fragments by combining 158 fragments of STR alleles obtained by nested PCR and two fragments artificially obtained from the sex-determination marker. By including the new allelic ladder in analysis of samples amplified with the Canine Kit, the accuracy and reliability of data analysis were improved. Application of this allelic ladder would be helpful for interlaboratory data sharing and standardization of canine genotype databases.


Subject(s)
Alleles , Dogs/genetics , Genotype , Genotyping Techniques , Animals , DNA Fingerprinting , Forensic Genetics/methods , Microsatellite Repeats , Polymerase Chain Reaction , Reproducibility of Results
11.
Healthcare (Basel) ; 9(6)2021 May 29.
Article in English | MEDLINE | ID: mdl-34072444

ABSTRACT

Oral breathing directly affects behavioral performance and dental health. Various relationships between oral breathing and periodontal disease have been well-described. However, the effect of oral breathing on cognitive performance remains unclear. This study aimed to investigate the effects of oral breathing on cognitive function using functional magnetic resonance imaging (fMRI). Twenty-two healthy participants (mean age, 22.27 ± 1.42 years) performed a two-back (2B) working memory fMRI task using a 3T MRI scanner while breathing through their oral or nasal passage. Functional activity analysis was performed using a statistical parametric mapping software package. One-sample group analyses were performed in 2B > Rest contrast. Functional connectivity analysis was conducted using MATLAB-based imaging software. Mixed ANOVA analysis was performed. The results showed more brain activation and connection during nasal breathing than during oral breathing. For Nasal > Oral contrast, various functional connections are known to have a significant relationship with working memory, including the left cerebellum, left and right inferior parietal gyrus. This can be significant evidence to demonstrate that oral breathing is an inappropriate method for intellectual activity using brain imaging techniques. Therefore, this study suggests that changing various habits related to oral breathing is important for cognitive function.

12.
Leg Med (Tokyo) ; 52: 101902, 2021 Sep.
Article in English | MEDLINE | ID: mdl-33964678

ABSTRACT

Casework evidence samples are likely to be placed under diverse and harsh environments as compared to quantified DNA samples including serial-diluted standard DNA samples. Internal validation of a novel STR kit using casework evidence sample, which is conducted according to various conditions such as DNA contamination and degradation, is crucial before being used as a forensic application. Therefore, this study aimed to elucidate the reliability of the Investigator® 24plex QS kit through DNA derived from casework evidence and to assess whether it is applicable to STR analysis together with PowerPlex® Fusion System and GlobalFiler™ PCR Amplification Kit. DNA was extracted from 189 casework evidence samples in a total of 77 cases. The mismatch of the allelic size of this kit through allelic sizing precision test, was suitable according to ENFSI guidelines. All heterozygous balance of the three kits were above 0.6 recommended value of ENFSI guideline. The number of allele drop-in was most frequent in the GlobalFiler™ PCR Amplification Kit. In addition, the number of allele drop-out was most frequent in the Investigator® 24plex QS kit. The cutoff concentration of DNA detected in three kits of one complete STR was approximately 45 pg/µL on average. Despite of several limitations, the Investigator® 24plex QS kit is considered to have the capability to be used for STR analysis of casework evidence samples.


Subject(s)
Forensic Genetics/methods , DNA/genetics , DNA Fingerprinting , Humans , Microsatellite Repeats/genetics , Polymerase Chain Reaction , Reproducibility of Results
13.
Brain Sci ; 10(10)2020 Oct 03.
Article in English | MEDLINE | ID: mdl-33022977

ABSTRACT

The problems of mouth breathing have been well-studied, but the neural correlates of functional connectivity (FC) still remain unclear. We examined the difference in FC between the two types of breathing. For our study, 21 healthy subjects performed voluntary mouth and nasal breathing conditions during a resting state functional magnetic resonance imaging (fMRI). The region of interest (ROI) analysis of FC in fMRI was conducted using a MATLAB-based imaging software. The resulting analysis showed that mouth breathing had widespread connections and more left lateralization. Left inferior temporal gyrus had the most left lateralized connections in mouth breathing condition. Furthermore, the central opercular cortex FC showed a significant relationship with mouth breathing. For nasal breathing, the sensorimotor area had symmetry FC pattern. These findings suggest that various FCs difference appeared between two breathing conditions. The impacts of these differences need to be more investigated to find out potential link with cognitive decline in mouth breathing syndrome.

14.
J Forensic Sci ; 65(6): 2117-2120, 2020 Nov.
Article in English | MEDLINE | ID: mdl-32725909

ABSTRACT

Lophophora is a member of the Cactaceae family, which contains two species: Lophophora williamsii and L. diffusa. Lophophora williamsii is an illegal plant containing mescaline, a hallucinogenic alkaloid. In this study, a novel method based on a single nucleotide polymorphism (SNP) assay was developed for identifying L. williamsii; this assay reliably detects SNPs within chloroplast DNA (rbcL, matK, and trnL-trnF IGS) and was validated for identifying Lophophora and L. williamsii simultaneously. The chloroplast DNA sequences from four L. williamsii and three L. diffusa plants were obtained and compared using DNA sequence data from approximately 300 other Cactaceae species available in GenBank. From this sequence data, a total of seven SNPs were determined to be suitable for identifying L. williamsii. A multiplex assay was constructed using the ABI PRISM® SNaPshot™ Multiplex Kit (Applied Biosystems, Forster City, CA) to analyze species-specific SNPs. Using this multiplex assay, we clearly distinguished the Lophophora among 19 species in the Cactaceae family. Additionally, L. williamsii was distinguished from L. diffusa. These results suggest that the newly developed assay may help resolve crimes related to illegal distribution and use. This multiplex assay will be useful for the genetic identification of L. williamsii and can complement conventional methods of detecting mescaline.


Subject(s)
Cactaceae/genetics , DNA, Chloroplast/genetics , Polymorphism, Single Nucleotide , Cactaceae/chemistry , Forensic Genetics/methods , Humans , Illicit Drugs , Mescaline/analysis , Multiplex Polymerase Chain Reaction , Sequence Analysis, DNA
15.
Ergonomics ; 63(11): 1359-1370, 2020 Nov.
Article in English | MEDLINE | ID: mdl-32552557

ABSTRACT

While working is more comfortable in a supine position and healthier in a standing, most people work in a sitting. However, it is unclear whether there are differences in brain activity efficiency in different postures. Here, we, therefore, compared changes in brain activity across three different postures to determine the optimal posture for performing working memory tasks. Their effect on brain activity was examined using EEG signals together with the information of accuracy and reaction times during 2-back task in 24 subjects. Substantial differences in brain waves were observed at sitting and standing positions compared to the supine, especially in delta waves and frontal lobe, where is known to improve the modulation of brain activity efficiently. Brain efficiency was higher during standing and sitting than in a supine. These findings show that postural changes may affect the efficiency of brain activity during working memory tasks. Practitioner summary: Differences in brain efficiency between different postures during working memory tasks have not been explored. This study suggests that efficiency in several brain areas is higher during sitting and standing than in a supine position. This finding has important implications regarding workplace environments. Furthermore, this result would be useful to improve accomplishment and reduce negative effects of work posture. Abbreviations: EEG: electroencephalogram; PSQI: Pittsburgh sleep quality index; KSS: Karolinska sleepiness scale; FFT: fast fourier transform; ROI: region of interest; ANS: autonomic nervous system; Fp: prefrontal; AF: anterior frontal; frontal; Fz: midline frontal; temporal; central; Cz: midline central; P: parietal; Pz: midline parietal; O: occipital; Oz: midline occipital.


Subject(s)
Brain/physiology , Memory, Short-Term/physiology , Posture/physiology , Adult , Electroencephalography , Female , Humans , Male , Occupational Health , Surveys and Questionnaires , Task Performance and Analysis , Workplace , Young Adult
16.
J Dermatol Sci ; 98(2): 128-136, 2020 May.
Article in English | MEDLINE | ID: mdl-32354609

ABSTRACT

BACKGROUND: Autophagy is a catabolic process for eliminating damaged organelles or proteins to maintain cellular homeostasis. Recently, lipids have been demonstrated to be targets for autophagosomal degradation. Therefore, autophagy might be involved in sebaceous gland homeostasis, however, relevant data are lacking. OBJECTIVES: We investigated the role of autophagy in sebaceous lipogenesis and its regulatory mechanisms in human SZ95 sebocytes. We also examined the possible role of autophagy in 13-cis-retinoic acid (13-cis-RA)-mediated sebosuppression. METHODS: Autophagy markers expression was examined by immunohistochemistry in normal and acne lesional skin. SZ95 sebocytes were treated with autophagy inhibitors under starvation or treated with a combination of testosterone and linoleic acid (testosterone/LA), with or without autophagy inducer rapamycin or 13-cis-RA. Lipids were assessed by BODIPY and quantitative Nile Red staining. Autophagy-related gene 7 small interference RNA was used to confirm the role of autophagy on the sebosuppressive effect of rapamycin or 13-cis-RA. RESULTS: Autophagy markers were strongly expressed in the maturing sebaceous gland cells in healthy skin, whereas downregulated in the acne-involved sebaceous glands. Testosterone/LA or insulin-like growth factor-1 inhibited starvation-induced sebocyte autophagy. Pharmacological inhibition of autophagy led to increased sebaceous lipid accumulation. Contrary, rapamycin inhibited the testosterone/LA-induced lipogenesis and expression of fatty acid synthesis genes via activating the autophagy pathway. 13-cis-RA increased autophagy in SZ95 sebocytes, partly via FoxO1 activation, and inhibition of autophagy abolished the sebosuppressive effect of 13-cis-RA. CONCLUSIONS: Autophagy plays an important role in the modulation of lipogenesis in human sebocytes and is involved in the sebostatic effect of 13-cis-RA.


Subject(s)
Acne Vulgaris/drug therapy , Autophagy/drug effects , Isotretinoin/pharmacology , Sebaceous Glands/drug effects , Acne Vulgaris/pathology , Autophagy/physiology , Cell Line , Humans , Isotretinoin/therapeutic use , Lipogenesis/drug effects , Lipogenesis/physiology , Sebaceous Glands/cytology , Sebaceous Glands/pathology , Sirolimus/pharmacology
17.
Int J Legal Med ; 134(4): 1551, 2020 07.
Article in English | MEDLINE | ID: mdl-32219527

ABSTRACT

The above article was published with two author names being incorrect. The published paper states "'Hyeun Kyu Yoon and Ki min Seong", whereas it should be "'Hyun Kyu Yoon and Ki Min Seong".

18.
Int J Legal Med ; 134(3): 963-975, 2020 05.
Article in English | MEDLINE | ID: mdl-31418076

ABSTRACT

DNA quantification is an essential step for successful multiplex short tandem repeat (STR) polymerase chain reactions (PCR), which are used for confirming identities using human genomic DNA. The new DNA quantification kit, named the National Forensic Service Quantification (NFSQ) kit, simultaneously provides total human DNA concentration, human male DNA concentration, and a DNA degradation index (DI) using multiplex TaqMan fluorescent probes. The NFSQ was validated according to developmental validation guidelines from the SWGDAM and MIQE. NFSQ detected up to 0.00128 ng/µL and could detect male DNA up to a 1:8000 ratio of male to female DNA. In PCR inhibitor tests, NFSQ could measure DNA at a concentration of 200 ng/µL of humic acid and 600 µM of hematin. The NFSQ kit showed a DI value trend similar to other qPCR kits. In the reproducibility study, the coefficient of variation of the NFSQ kit was within 10%. The quantitative results of the casework samples obtained using the NFSQ kit were consistent with the STR interpretation results. The NFSQ kit can be useful in the human identification process, as it has detection capabilities similar to those of other comparable quantification kits.


Subject(s)
DNA Fingerprinting/instrumentation , Sequence Analysis, DNA/methods , Animals , Female , Fluorescent Dyes , Genetic Markers , Humans , Male , Microsatellite Repeats , Nucleic Acid Denaturation , Real-Time Polymerase Chain Reaction , Reproducibility of Results , Republic of Korea , Sensitivity and Specificity , Species Specificity
19.
J Forensic Sci ; 65(1): 209-213, 2020 Jan.
Article in English | MEDLINE | ID: mdl-31433497

ABSTRACT

When using non-FTA cards in commercial multiplex STR kits for direct PCR, pretreatment steps with specific buffers are recommended. Here, we designed a rapid direct PCR method utilizing a non-FTA card, Oral Cell Sampling Kit, by omitting the pretreatment step involving Prep-n-Go™ Buffer, and it showed compatibility with the GlobalFiler™ Express PCR Amplification Kit, GlobalFiler™ PCR Amplification Kit, and PowerPlex® Fusion system. To optimize the PCR conditions, we tested the method with different final PCR volumes and cycles. Finally, we conducted a performance test using 50 Korean buccal samples and confirmed the high performance of the method, detecting more than 90% of the samples with full profiles when using GlobalFiler™ PCR Amplification Kit and PowerPlex® Fusion system at 29 cycles in a 10 µL final PCR volume. Thus, we report a simple direct PCR set-up to analyze reference samples collected using a non-FTA card manufactured in Korea.


Subject(s)
Polymerase Chain Reaction/methods , Specimen Handling/instrumentation , DNA Fingerprinting/methods , Female , Humans , Microsatellite Repeats , Mouth Mucosa/chemistry , Republic of Korea , Specimen Handling/methods
20.
Int J Legal Med ; 133(6): 1711-1719, 2019 Nov.
Article in English | MEDLINE | ID: mdl-31388795

ABSTRACT

Inference of ancestry from biological evidence can provide investigative information, especially for unknown DNA donors. Although tools for predicting ancestry have been developing, ancestry research focusing on populations relevant for South Korea is not common and markers are seldom chosen specifically to differentiate Koreans from other East Asian and South East Asian populations. Here, we report ancestry informative markers (AIMs) for distinguishing six East/South East Asian regional populations: China, Japan, Indonesia, Philippines, South Korea and Thailand. Individual genotypes from these six populations were available in PanSNPdb: The HUGO Pan-Asian SNP Database. To select AIMs, we calculated four population divergence metrics for each SNP: Nei's FST, Rosenberg's Informativeness (In), the average absolute allele frequency difference between populations (δFmean) and the maximum allele frequency difference between populations (δFmax). Based on these values, we selected 100 single nucleotide polymorphisms (SNPs) for distinguishing the six populations, 13 of which exhibited large allele frequency differences between Koreans and non-Koreans. To assess the performance of the AIMs, we performed principal coordinates analysis (PCoA) on the individuals from all six populations and inferred ancestral population clusters using the STRUCTURE program. In conclusion, we found that the selected AIMs can be applied to distinguish the six East/South East Asian groups and we suggest the markers in this study will be helpful to establish ancestry panels for Korea and neighbouring populations.


Subject(s)
Asian People/genetics , Genetic Markers , Genetics, Population , Polymorphism, Single Nucleotide , Asia , DNA Fingerprinting , Databases, Genetic , Gene Frequency , Genotype , Humans , Principal Component Analysis
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