scholarly journals Development and validation of a multiplex 19 X-chromosomal short tandem repeats typing system for forensic purposes

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Juan Jia ◽  
Xu Liu ◽  
Qingwei Fan ◽  
Chen Fang ◽  
Mengchun Wang ◽  
...  
Keyword(s):  
Tandem Repeats ◽  
Dna Analysis ◽  
Forensic Application ◽  
Str Typing ◽  
Sex Typing ◽  
Typing System ◽  
Development And Validation ◽  
Multiplex System ◽  
Short Tandem ◽  
Forensic Parameters

AbstractX-chromosome short tandem repeat (X-STR) markers are a powerful complementary system used for paternity and forensic casework. This study presents the development and validation of a new highly efficient multiplex-fluorescent-labeled 19 X-STR typing system, including DXS10079, DXS101, DXS10135, DXS10162, DXS6795, DXS6800, DXS6803, DXS6807, DXS6809, DXS6810, DXS7133, DXS7423, DXS981, DXS9902, DXS9907, GATA165B12, GATA172D05, GATA31E08 and HPRTB along with sex-typing locus, amelogenin. The system was validated according to guidelines issued by the Scientific Working Group on DNA Analysis Methods. Allele frequency and forensic parameters were investigated from 1085 (494 males and 591 females) unrelated Beijing Han individuals, the combined power of discrimination by the 19 X-STR loci in females and males, as well as the combined mean exclusion chance in trios and duos, were 0.999999999999999995, 0.99999999995, 0.9999999995, and 0.9999996, respectively. The results demonstrate that this multiplex system is robust and reliable, and considered to be a powerful tool for forensic application.

Development and validation of a novel 29-plex Y-STR typing system for forensic application

2020 ◽  
Vol 44 ◽  
pp. 102169 ◽  
Author(s):  
Min Li ◽  
Wei Zhou ◽  
Yilun Zhang ◽  
Lei Huang ◽  
Xinjie Wang ◽  
...  
Keyword(s):  
Forensic Application ◽  
Str Typing ◽  
Typing System ◽  
Development And Validation

scholarly journals Identification and Characterization of Nine Novel X-Chromosomal Short Tandem Repeats on Xp21.1, Xq21.31, and Xq23 Regions

2021 ◽  
Vol 12 ◽  
Author(s):  
Qinrui Yang ◽  
Jinglei Qian ◽  
Chengchen Shao ◽  
Yining Yao ◽  
Zhihan Zhou ◽  
...  
Keyword(s):  
Short Tandem Repeats ◽  
Tandem Repeats ◽  
Mapping Function ◽  
Str Analysis ◽  
The Family ◽  
Further Development ◽  
Multiplex System ◽  
Short Tandem ◽  
Forensic Parameters

The application of X-chromosomal short tandem repeats (X-STRs) has been recognized as a powerful tool in complex kinship testing. To support further development of X-STR analysis in forensic use, we identified nine novel X-STRs, which could be clustered into three linkage groups on Xp21.1, Xq21.31, and Xq23. A multiplex PCR system was built based on the electrophoresis. A total of 198 unrelated Shanghai Han samples along with 168 samples from 43 families was collected to investigate the genetic polymorphism and forensic parameters of the nine loci. Allele numbers ranged from 5 to 12, and amplicon sizes ranged from 146 to 477 bp. The multiplex showed high values for the combined power of discrimination (0.99997977 in males and 0.99999999 in females) and combined mean exclusion chances (0.99997918 and 0.99997821 in trios, 0.99984939 in duos, and 0.99984200 in deficiency cases). The linkage between all pairs of loci was estimated via Kosambi mapping function and linkage disequilibrium test, and further investigated through the family study. The data from 43 families strongly demonstrated an independent transmission between LGs and a tight linkage among loci within the same LG. All these results support that the newly described X-STRs and the multiplex system are highly promising for further forensic use.

scholarly journals Genetic analysis of DXS101and DXS6807 short tandem repeat (X-STR) in samples of Iraqi Arab male population

2020 ◽  
Vol 14 (1) ◽  
pp. 34-38
Author(s):  
Hayder Allawi Khaleefah ◽  
Salwa Jaber Abdullah Al-Awadi ◽  
Zaid Nsaif Abbas Al-Tameemi
Keyword(s):  
Information Content ◽  
Polymorphism Information Content ◽  
Tandem Repeats ◽  
Forensic Application ◽  
Male Population ◽  
Forensic Efficiency ◽  
Back Ground ◽  
Forensic Markers ◽  
Short Tandem ◽  
Apparently Healthy

Back ground: X-chromosomal short tandem repeats (X-STRs) have assured to be informative and particular role in complex relationship testing. DXS6807 known as tetra nucleotides polymorphism representing eight alleles of 251-275 bp in length. DXS6807 is located in, at XP 22.2, at a genetic distance of more than 87 and 151 Cm of X-chromosome. DXS101 is located104.9–121 cM from the Xp-telomere (Xp-tel) corresponding to cytogenetic position in Xq21.33–Xq22.3.  Objective: The aim of this present study   investigates the allele frequency of two markers DXS101, DXS6807 and forensic efficiency parameters for sample of Arabic Iraqi males. Material and methods: The population of this study includes 200 males apparently healthy unrelated participants from different region of Baghdad city, their ages ranged between (20-50) years. The Genomic DNA extracted and purified successfully from blood samples.  Results: The forensic efficiency parameters result for these markers were following: polymorphism information content (PIC) of 0.834708, power of discrimination (PD) in male 0.851750, Power of exclusion (PE) 0.698316, MEC Krüger0.511679, MEC Kishida 0.694890. The forensic efficiency parameters analyzing from Arabic population were Power of discrimination (PD) = 0.73405, Polymorphism information content (PIC) =0.69489, Power of exclusion (PE) =0.482879.MEC Krüger =0.511679, MEC Kishida = 0.694890. Conclusions: The information provided establish this X-linked microsatellite marker as a valuable strategy for forensic application. DXS101is and DXS6807 recently consider more stable and suitable forensic markers for forensic application.

scholarly journals Validation of a 6-Dye Short Tandem Repeat System: A Dry Kit With Lyophilized Amplification Reagent

2021 ◽  
Vol 12 ◽  
Author(s):  
Shuanglin Li ◽  
Jinfeng Lin ◽  
Honglei Hao ◽  
Haiying Jin ◽  
Danlu Song ◽  
...  
Keyword(s):  
Tandem Repeats ◽  
Dna Analysis ◽  
Cumulative Probability ◽  
Optimal Range ◽  
System A ◽  
Total Input ◽  
Amplification System ◽  
Autosomal Short Tandem Repeats ◽  
The Stability ◽  
Short Tandem

The SureID®S6 system used a lyophilized pellet as the amplification reagent to enable multiplexing of sex-determining marker Amelogenin, 21 autosomal short tandem repeats (STRs), and one Y-STR. To assess the performance, reliability, and limitation of the dry amplification system, the validation studies including PCR condition, reproducibility, sizing and precision, analytical threshold calculation, sensitivity and stochastic threshold calculation, species specificity, stability, mixture, case sample, and population and concordance were conducted according to the Scientific Working Group on DNA Analysis Methods (SWGDAM) Validation Guidelines. Experimental data suggested that the optimal range of total input DNA was from 125 to 500 pg; the appropriate analytical threshold was 80 relative fluorescence units (RFUs) while the stochastic threshold was 260 RFUs; for the stability studies, SureID®S6 system could resist against less than 500 μmol/L of hematin, 100 ng/μl of humic acid, 4 mM of indigotin, 800 mM of tannic acid, and 800 mM of calcium ion. Population and concordance studies using 500 unrelated individuals showed that the combined probability of discrimination (CPD) and cumulative probability of exclusion (CPE) values were 0.999999999999 and 0.999999998416, respectively. The genotypes for the same sample were concordant with the previously validated HUAXIA™ Platinum kit. The validation results demonstrated that the SureID®S6 system could be used for forensic applifications.

Generation and characterization of human Fetal membrane and Decidual cell lines for reproductive biology experiments

2021 ◽  
Author(s):  
Enkhtuya Radnaa ◽  
Rheanna Urrabaz-Garza ◽  
Nathan D Elrod ◽  
Mariana Castro Silva ◽  
Richard Pyles ◽  
...  
Keyword(s):  
Cell Lines ◽  
Short Tandem Repeats ◽  
Tandem Repeats ◽  
Dna Analysis ◽  
Fetal Membrane ◽  
Primary Cells ◽  
Immortalized Cells ◽  
Passage Number ◽  
Maternal Decidua ◽  
Short Tandem

Abstract Human fetal membrane and maternal decidua parietalis form one of the major feto-maternal interfaces during pregnancy. Studies on this feto-maternal interface is limited as several investigators have limited access to the placenta, and experience difficulties to isolate and maintain primary cells. Many cell lines that are currently available do not have the characteristics or properties of their primary cells of origin. Therefore, we created, characterized the immortalized cells from primary isolates from fetal membrane-derived amnion epithelial cells, amnion and chorion mesenchymal cells, chorion trophoblast cells and maternal decidua parietalis cells. Primary cells were isolated from a healthy full-term, not in labor placenta. Primary cells were immortalized using either a HPV16E6E7 retroviral or a SV40T lentiviral system. The immortalized cells were characterized for the morphology, cell type-specific markers, and cell signalling pathway activation. Genomic stability of these cells was tested using RNA seq, karyotyping, and short tandem repeats DNA analysis. Immortalized cells show their characteristic morphology, and express respective epithelial, mesenchymal and decidual markers similar to that of primary cells. Gene expression of immortalized and primary cells were highly correlated (R = 0.798 to R = 0.974). Short tandem repeats DNA analysis showed in the late passage number (>P30) of cell lines matched 84-100% to the early passage number (<P10) of the cell lines revealing there were no genetic drift over the passages. Karyotyping also revealed no chromosomal anomalies. Creation of these cell lines can standardize experimental approaches, eliminate subject to subject variabilities, and benefit the reproductive biological studies on pregnancies by using these cells.

Multiplex fluorescent analysis of four short tandem repeats for rapid haemophilia A molecular diagnosis

2005 ◽  
Vol 94 (11) ◽  
pp. 1099-1103 ◽  
Author(s):  
Jorge Sánchez-García ◽  
Dominique Gallardo ◽  
Lorena Ramírez ◽  
Francisco Vidal
Keyword(s):  
Molecular Diagnosis ◽  
Short Tandem Repeats ◽  
Tandem Repeats ◽  
Dna Analysis ◽  
Single Cells ◽  
Genetic Diagnosis ◽  
Automated Method ◽  
Fluorescent Method ◽  
Fluorescent Pcr ◽  
Short Tandem

SummaryIndirect molecular diagnosis of hemophilia A (HA) is carried out by analyzing intragenic polymorphic markers described along the coagulation factorVIII (FVIII) gene. Several studies have demonstrated that the two commonly used intronic short tandem repeats (STR13 and STR22) located in the FVIII gene are highly informative for this task. Two extragenic markers closely linked to FVIII (DXS1073 and DXS1108) have also been described as valuable tools for gene tracking. The objective of the present work was to develop a rapid, single-tube automated method to simultaneously analyze these four STRs. Consistent amplification was achieved by quadruplex fluorescent PCR and the products were analyzed by capillary electrophoresis. Validation of the method included DNA analysis of 88 individuals from a control population, 45 HA patients and 32 individuals from 10 HA-affected families. Statistical study showed that the STR13, STR22 and DXS1108 loci were in significant linkage disequilibrium, whereas DXS1073 was not. Nevertheless, the combination of the four markers offered a high heterozygosity rate (>90%) that improved tracing of FVIII gene inheritance. Optimal results with application to single cells in a HA preimplantation genetic diagnosis (PGD) protocol demonstrated the sensitivity of the technique. In conclusion, the automated fluorescent method described is an extremely rapid, simple and highly informative one that is easy to standardize and allows direct comparison of results among different groups working with genetic counseling, prenatal diagnosis and PGD in HA-affected families.

Genetic analysis of 12 X-short tandem repeats loci in a northern Thai population

2020 ◽  
pp. 002580242096500
Author(s):  
Supakit Khacha-ananda ◽  
Phatcharin Mahawong
Keyword(s):  
Genetic Polymorphism ◽  
Allele Frequency ◽  
Short Tandem Repeats ◽  
Tandem Repeats ◽  
Thai Population ◽  
Weinberg Equilibrium ◽  
Hardy Weinberg Equilibrium ◽  
Forensic Genetic ◽  
Short Tandem ◽  
Forensic Parameters

Short tandem repeats (STRs) are widely used as DNA markers in paternity testing and criminal investigations because of their high genetic polymorphism among individuals in population. However, many factors influence genetic variations of STRs. Therefore, understanding STR information within individual populations could provide database and scientifically reliable STR genotyping for forensic genetic purposes. We aimed to examine allele frequencies of X-STRs, including some forensic parameters, in a northern Thai population. A retrospective descriptive study was conducted by collecting X-STR data from unrelated individuals living in a northern region of Thailand. The allele frequency and forensic parameters – for example polymorphism information content (PIC), power of discrimination in females and males (PDf and PDm), mean exclusion chance (MEC) and haplotype frequency – were calculated. The Hardy–Weinberg equilibrium was analysed. A total of 132 alleles were observed, with corresponding allele frequency ranging from 0.0064 to 0.4904. The PIC of all loci was >0.6, representing high genetic polymorphism, except DXS8378 and DXS7423. Notably, DXS10135 was the most diverse loci with the highest PD and MEC, while DXS7423 was the least polymorphic marker with the lowest PD and MEC. The highest haplotype diversity in male data was on linkage group III (DXS10101-DXS10103-HPRTB) by 0.9895. The genetic distance analysis demonstrated that the northern Thai population had a close relationship with Taiwanese (DA = 0.023). There are no significant deviations among the Hardy–Weinberg equilibrium except DXS10148. This study has established a northern Thai X-STRs reference database to be used as a tool for forensic genetic purposes.

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