Hungarian population data of eight X-linked markers in four linkage groups

The X chromosomal STR markers DXS10135 and DXS8378 in linkage group 1, DXS7132 and DXS10074 in linkage group 2, HPRTB and DXS10101 in linkage group 3, and DXS10134 and DXS7423 in linkage group 4 were studied in the Hungarian population. After genotyping unrelated men (219) and women (165), forensic efficiency parameters were calculated. Deviations from Hardy-Weinberg equilibrium could not be detected. There were several microvariant and rare alleles were sequenced: four in locus DXS10135 (alleles 17.1, 18.1, 20.1 and 26.1), one in locus DXS10074 (alleles 11), three in locus DXS10101 (alleles 26, 34.2 and 35) and five in locus DXS10134 (alleles 35.3, 37.2, 38.2, 39.2, 41).     

A LDR‐PCR Approach for Multiplex Polymorphisms Genotyping of Severely Degraded DNA with Fragment Sizes <100 bp*

Abstract: Reducing amplicon sizes has become a major strategy for analyzing degraded DNA typical of forensic samples. However, amplicon sizes in current mini‐short tandem repeat‐polymerase chain reaction (PCR) and mini‐sequencing assays are still not suitable for analysis of severely degraded DNA. In this study, we present a multiplex typing method that couples ligase detection reaction with PCR that can be used to identify single nucleotide polymorphisms and small‐scale insertion/deletions in a sample of severely fragmented DNA. This method adopts thermostable ligation for allele discrimination and subsequent PCR for signal enhancement. In this study, four polymorphic loci were used to assess the ability of this technique to discriminate alleles in an artificially degraded sample of DNA with fragment sizes <100 bp. Our results showed clear allelic discrimination of single or multiple loci, suggesting that this method might aid in the analysis of extremely degraded samples in which allelic drop out of larger fragments is observed.     

Texas Population Substructure and Its Impact on Estimating the Rarity of Y STR Haplotypes from DNA Evidence*

Abstract: Three sampled populations of unrelated males—African American, Caucasian, and Hispanic, all from Texas—were typed for 16 Y short tandem repeat (STR) markers using the AmpFlSTR^® Yfiler^TM kit. These samples also were typed previously for the 13 core CODIS autosomal STR loci. Most of the 16 marker haplotypes (2478 out of 2551 distinct haplotypes) were observed only once in the data sets. Haplotype diversities were 99.88%, 99.89%, and 99.87% for the African American, Caucasian, and Hispanic sample populations, respectively. F_ST values were very small when a haplotype comprised 10–16 markers. This suggests that inclusion of substructure correction is not required. However, haplotypes consisting of fewer loci may require the inclusion of F_ST corrections. The testing of independence of autosomal and Y STRs supports the proposition that the frequencies of autosomal and Y STR profiles can be combined using the product rule.     

Developmental Validation of the PrepFiler™ Forensic DNA Extraction Kit for Extraction of Genomic DNA from Biological Samples*

Abstract: The PrepFiler? Forensic DNA Extraction Kit enables isolation of genomic DNA from a variety of biological samples. The kit facilitates reversible binding of DNA with magnetic particles resulting in high DNA recovery from samples with very low and high quantities of biological materials: 0.1 and 40 μL of human blood (donor 2) provided 14 and 2883 ng of DNA, respectively. Following the revised SWGDAM guidelines, performance of the developed method was investigated using different sample types including saliva on swabs, semen stains on cotton fabric, samples exposed to environment, samples with polymerase chain reaction (PCR) inhibitors, blood stains (on denim, cotton cloth, and FTA^® paper), and touch evidence‐type samples. DNA yields for all samples tested were equal or better than those obtained by both phenol–chloroform extraction and commercial kits tested. DNA obtained from these samples was free of detectable PCR inhibitors. Short tandem repeat profiles were complete, conclusive, and devoid of PCR artifacts.     

A Cannabis sativa STR Genotype Database for Australian Seizures: Forensic Applications and Limitations*

Abstract: A genetic database was established with the aim of documenting the genetic diversity of Cannabis sativa in Australia for future utilization in forensic investigations. The database consisted of genotypes at 10 validated short tandem repeat loci for 510 plants representing drug seizures from across Australia and 57 fiber samples. A total of 106 alleles and 314 different genotypes were detected. All fiber samples exhibited unique genotypes while 55% of the drug samples shared a genotype with one or more samples. Shared genotypes were mostly found within seizures; however, some genotypes were found among seizures. Statistical analysis indicated that genotype sharing was a consequence of clonal propagation rather than a lack of genetic resolution. Thus, the finding of shared genotypes among seizures is likely due to either a common supplier, or direct links among seizures. Notwithstanding the potential intelligence information provided by genetic analysis of C. sativa, our database analysis also reveals some present limitations.     

甲醛固定石蜡包埋组织STR分型检测

目的评估10%甲醛固定石蜡包埋组织STR分型结果的影响因素。方法采用QIAGEN法、IQ法、Chelex法对2具新鲜尸体在尸检时常规制备的心、脑、肝、脾、肾、肺、胃、肠石蜡包埋组织进行DNA提取,用AmpFlSTR Identifiler试剂盒进行PCR扩增,在3100-Avant上完成片段分析。另外对15个案例中室温保存1～5年的心、肝、肺、肠存档石蜡包埋组织共56份采用同样的方法进行STR分型。以STR基因座检出率评估分型有效性。结果各种组织DNA片段均随着保存时间延长而持续降解,其中心、肺组织STR基因座检出率与保存时间存在线性相关。相同保存时间时,各种组织基因座检出率差异有统计学意义。其中以肺组织在不同保存时间中基因座检出率最高。结论在甲醛固定时间一定的条件下,存放时间、组织类型、DNA提取方法和PCR模板质量浓度是影响石蜡包埋组织STR分型的重要因素。     

Identification of gestational trophoblastic disease in a sexual assault case

In this study, gestational trophoblastic disease (GTD) was observed by short tandem repeat (STR) typing from the aborted tissues in a sexual assault case. By histological screening, the fetal tissue could not be distinguished from the maternal tissue in this case. Therefore, five specimens were collected randomly from the aborted tissues for DNA analysis. STR typing was performed by the commercial ABI Identifiler kit. The results showed that three specimens were of the maternal origin, one was a mixture of the mother and male fetus, and the other one was of male fetal origin with partial triploid. Three alleles were identified in each locus of D8S1179, D7S820 and VWA for the fetal specimen. For these three alleles, one matched the maternal origin and the others matched the putative paternal origin (suspect). Analysis of the Y-STR by using the commercial ABI Y-Filer kit, the fetal types matched the types of the suspect. We reported the case of partial mole on forensic evidence and gave the valuable information from its identification.     

亳州市84例新型冠状病毒肺炎的辨证论治分析

目的 分析安徽省亳州市84例新型冠状病毒肺炎患者的中医证候及辨证论治特点。方法 观察新型冠状病毒肺炎患者的临床症状、舌象,进行辨证分型和辨证治疗。结果 84例患者中,临床治疗期湿毒郁肺证54例,寒湿阻肺证26例,疫毒闭肺证4例,分别选用清肺排毒汤1、2、3号方治疗;恢复期肺脾气虚证68例,气阴两虚证16例,分别选用自拟康复1、2号方治疗。结论 亳州市确诊的新型冠状病毒肺炎患者临床治疗期主要中医证型为湿毒郁肺证（湿热并重）、寒湿阻肺证（寒微湿重）,疫毒闭肺证少见,恢复期肺脾气虚证多见,气阴两虚证少见。应根据国家卫生健康委员会制定的《新型冠状病毒肺炎诊疗方案（试行第六版）》和亳州市患者的证候特点进行辨证论治。     

Validation of the Investigator 24plex QS Kit: a 6-dye multiplex PCR assay for forensic application in the Chinese Han population

The Investigator 24plex QS Kit (QIAGEN, Hilden, Germany) is a 6-dye fluorescent chemistry short tandem repeat (STR) polymerase chain reaction (PCR) amplification system that simultaneously amplifies 20 of the expanded Combined DNA Index System (CODIS) core STR loci, SE33, DYS391, and the standard sex-determining locus, amelogenin, as well as two special internal performance quality sensor controls (QS1 and QS2), which are included in the primer mix to check the PCR performance. This study was designed to be a pilot evaluation of this STR-PCR kit in a Chinese Han population regarding the PCR conditions, sensitivity, precision, accuracy, repeatability, reproducibility, and concordance; tolerance to PCR inhibitors; applicability to real “forensic-type” samples; species specificity; mixture, balance and stutter analyses, and utility in a population investigation. The exhaustive validation studies demonstrated that the Investigator 24plex QS system is accurate, sensitive and robust for STR genotyping. In addition, these genetic markers in the population data in our study indicated that they can also be useful for forensic identification and paternity testing in the Chinese Han population.     

Forensic examination and application of areca nuts as material evidence

In this study, we aimed to explore the possibility of DNA analysis of areca nut as material evidence and the value of short tandem repeat (STR) typing of areca nut as material evidence under the condition of simulating external environment. In this study, water soaking, soil burial, sun exposure, and wet environment were used to treat areca nut residues. Chelex 100 was used to extract DNA, the PowerPlex21 kit to amplify, and the ABI PRISM^® 310 Genetic Analyzer to analyze the DNA of areca nut residues. DNA and STR typing were performed to analyze the residue after chewing. The results showed that the number of residual sites decreased with time under the conditions of water soaking, soil burial, sun exposure, and wet environment. Thus, areca nut can be used as forensic material evidence for DNA analysis and individual identification.