Mitochondrial DNA regions HVI and HVII population data.

Data from 1393 unrelated individuals have been compiled from eight population groups: African Americans, Africans (Sierra Leone), U.S. Caucasians, Austrians, French, Hispanics, Japanese, and Asian Americans. The majority of the mtDNA sequences were observed only once within each population group (i.e., ranging from a low of 60.3% (35/58) of the Asian American sequences to a high of 85.3% (93/109) of the French sequences). Genetic diversity ranged from 0.990 in the African sample to 0.998 in African Americans. Random match probability ranged from 2.50% in the Asian American sample to 0.52% in U.S. Caucasians. The average number of nucleotide differences between individuals in a database is greatest for the African American and African samples (14.1 and 13.1, respectively), and the least variable are the Caucasians (ranging from 7.2 to 8.4). Substitutions are the predominate polymorphism, and at least 92% of the substitutions are transitions. The most prevalent transversions are As substituted for Cs and Cs substituted for As. For most population groups these transversions occurred predominately in the HVI region; however, the African, African American, and Hispanic samples also demonstrated a large portion of their C to A and A to C transversions in the HVII region (at sites 186 and/or 189). Most insertions occur in the HVII region at sites 309.1 and 315.1, within a stretch of C's. Insertions of an additional C are common in all population groups. The sequence data were converted to SSO mtDNA types and compared with population data on Caucasians, Africans, Asians, Japanese, and Mexicans described by Stoneking et al. [M. Stoneking, D. Hedgecock, R.G. Higuchi, L. Vigilant, H.A. Erlich, Population variation of human mtDNA control region sequences detected by enzymatic amplification and sequence-specific oligonucleotide probes, Am. J. Hum. Genet. 48 (1991) 370-382] using an R x C contingency table test. Differences between major population groups (i.e., between African, Caucasian, and Asian) are quite evident, and similar ethnic population groups carried similar SSO polymorphism frequencies. There were only a few SSO types that showed significant differences between subpopulation groups. The SSO data alone can not be used to describe the population genetics with complete sequence data. However, the results of the SSO comparisons are similar to other analyses, and differences in sequence data in regions HVI and HVII are greater between major population groups than between subgroups.     

线粒体DNA cytb和HVI的分析用于种属鉴定

目的以线粒体DNA为目标序列,探讨生物检材的种属来源问题。方法复合扩增线粒体DNA细胞色素b基因(Cb)片段和D-环HVI上人源特异性DNA片段,2%琼脂糖凝胶电泳检测复合扩增产物谱带;用常规测序技术获得种属来源不明的检材Cytb基因序列,登陆美国国家生物信息中心网站主页(http://www.ncbi.nlm.nih.gov),将Cytb序列的测序结果用BLAST2.2.9[2004.5.1]进行匹配查询,查询数据库中存在的与其相匹配的物种条目。结果检材经复合扩增后电泳检测可区分人源性检材和非人源性检材;用生物信息法可确定检材种属来源结论检测线粒体DNA细胞色素b基因和D-环HVI的有关序列可在DNA分子水平上鉴别人源性检材和非人源性检材,结合测序的分子生物信息学方法,可对检材进行种属鉴定。     

Optimization of a duplex amplification and sequencing strategy for the HVI/HVII regions of human mitochondrial DNA for forensic casework

A duplex primer set for the amplification of mitochondrial DNA HVI and HVII control regions was evaluated for the optimization of a DNA sequencing protocol suitable for forensic casework. HVI and HVII products, with the absence of non-specific products, could be detected by agarose gel electrophoresis when as little as 0.5 and 0.1pg of DNA were amplified for 34 and 38 cycles, respectively. Because HVI and HVII amplicons are co-synthesized in the duplex PCR, fewer steps are required (lessening the risk of cross contamination events) and more frugal use of precious extracted DNA samples is possible, both desirable features for forensic casework. The ABI Prism BigDyetrade mark version 1.1 chemistry provided high quality sequencing data, with little or no background noise and uniform peak heights, outcomes that favored reliable detection of heteroplasmy, particularly at early sequence reads (<40 bases). Optimal compromise between sensitivity and sequence accuracy in the absence of noise was achieved starting at 150 mitochondrial genome copies. The protocol is effective (no sequence errors) with highly degraded DNA (average detectable template size of 200bp). Dual artificial template mixtures with the minor component at 15% suggests that heteroplasmy should be detected at this level with confidence.     

Mitochondrial DNA control region variation in a population sample from Hong Kong, China

Entire mitochondrial control region sequences were generated from 377 unrelated individuals from urban Hong Kong. In line with other control region datasets from China, the sample from Hong Kong exhibited significant genetic diversity that was reflected in a random match probability of 0.19% and a mean pairwise difference of 13.14. A total of 305 haplotypes were identified, of which 262 were unique. These sequences will be made publicly available to serve as forensic mtDNA reference data for China.     

Mitochondrial DNA control region population data from Macedonia

Mitochondrial DNA sequences of the entire control region were analyzed in 200 unrelated individuals from Macedonia. A total of 163 different haplotypes were found as determined by 177 polymorphic sites. The probability of a random match was calculated as 1:121 (0.83%). The basic phylogenetic structure of the Macedonian population as derived from its haplogroup distribution is in agreement with other West-Eurasian populations. Upon publication, the population data are going to be available in the EMPOP database (www.empop.org) [W. Parson, A. Dür, EMPOP—a forensic mtDNA database, FSI:Genetics 1 (2) (2007) 88–92; W. Parson, A. Brandstätter, A. Alonso, N. Brandt, B. Brinkmann, A. Carracedo, et al., The EDNAP mitochondrial DNA population database (EMPOP) collaborative exercises: organisation, results and perspectives, Forensic Sci. Int. 139 (2–3) (2004) 215–226.].     

北京汉族群体线粒体DNA高变区的多样性

应用ＰＣＲ产物直接测序法研究北京汉族 10 0名无关个体线粒体ＤＮＡ序列特征 ,并对其进行统计学分析 ,现报道如下。1　材料与方法1 1　材料1 1 1　样本　无关个体血液样本 10 0份 ,取自本实验室日常办案的检材。1 1 2　引物　对ＨＶⅠ、ＨＶⅡ及其周围区域 ,根据Ａｎｄｅｒｓｏｎ[1] 序列 ,并参考文献 [2 ]自行设计引物。引物序列 (由ＧＩＢＩＣＯＬ公司合成 )为 :ｍｔ 1Ｈ5′ ＧＡＡＴＣＧＧＡＧＧＡＣＡＡＣＣＡＧＴＡＡＧ 3′Ｌ 5′ ＴＡＧＣＧＧＴＴＡＴＴＡＴＡＧＧＧＴ 3′ ;ｍｔ 0Ｈ5′ ＣＡＴＧＧＧＧＡＡＧＣＡＧＡＴＴＴＧ 3′…     

Metacarpal sexual determination in a Spanish population

Anthropologists and forensic pathologist determine the sex of skeletons by analyzing quantitative and qualitative characters in the bone remains. Generally, the skull and os coxae are the elements most used, but they are not always preserved. In such cases, the investigator needs to have available other techniques based on different remains. The aim of the present work is to develop and describe discriminating functions for sex determination in a recent Spanish population using metacarpal morphology. A sample of bones corresponding to a contemporary Spanish population deposited at the Complutense University of Madrid (UCM) was analyzed. This sample comprised 697 metacarpals, corresponding to 79 adult individuals (37 men and 42 women). These allowed us to obtain 120 unifactorial discriminant functions. We selected the 10 equations, one for each metacarpal from both hands, that provided the best sexual discrimination. The correct sex classification rank progressed from 81%, for right (R) metacarpals IV and V, to 91%, for left (L) metacarpal II. The results suggest that metacarpals are structures that can be used for sex determination in paleoanthropological and forensic identifications.     

Mitochondrial DNA population data of HVS-I and HVS-II sequences from a northeast German sample

Mitochondrial DNA sequences of the control region's two hypervariable regions HVS-I and HVS-II were determined for 213 unrelated west Eurasian individuals from northeast Germany (Mecklenburg). A total of 174 different mtDNA haplotypes were found, 25 of which were shared by more than 1 individual. The most frequent haplotypes were 263G-309.1C-315.1C, found in seven individuals, 263G-309.1C-309.2C-315.1C, found in six individuals and 263G-315.1C, found in five individuals. These sequences are also the most common haplotypes in other published European data sets. The sequence polymorphisms consisting of 150 polymorphic nucleotide positions were compared with other European databases. The genetic diversity and random match probability were calculated. Our results corroborate certain features which are characteristic for west Eurasian mtDNA population samples.     

Y-chromosome variation in a Norwegian population sample

Y-chromosome DNA profiles are promising tools in population genetics and forensic science. Here we present DNA profiles of 300 unrelated Y-chromosomes of Norwegian origin. The profile is composed of eight short tandem repeats (STRs) and one single nucleotide polymorphism (SNP). In more than 2/3 of the haplotypes the modular structure in the 5' end of the minisatellite locus DYF155S1 was revealed by minisatellite variant repeat PCR (MVR-PCR) These haplotypes were also typed for deletions of fragment 50f2C (DYF155S2). Allele distribution and paternity exclusion parameters are given for each marker. The degree of haplotype diversity and its implication for statistics are evaluated. In the 300 samples 177 different haplotypes were encountered, of which 137 were observed once only. Analysis showed that the main source of variation is within the population. The Fst values were less than 0.015 in general. Haplotype grouping by the SNP demonstrated two haplogroups (Tat/T and Tat/C). Haplogroup Tat/C--found in 5.7% of the present material - is the same haplogroup as encountered in 60% of Finnish males [Am. J. Hum. Genet. 62 (1998) 1171]. Mutation analysis in 150 father/son pairs (a total of 1200 meiotic events) revealed an average mutation frequency of 0.0042 (95% CI 0.0014-0.0097).     

Genetic data of 10 X-STRs in a Spanish population sample

In this work, we present population genetic data of 10 X-chromosome STRs (DXS8378, DXS9898, DXS8377, HPRTB, GATA172D05, DXS7423, DXS6809, DXS7132, DXS101 and DXS6789) obtained from sample of 145 unrelated female individuals belonging to Valencia (Spain), a region located in the east of the Iberian Peninsula. All the markers studied present high genetic diversities, similar to those previously reported in other European population samples. No deviations from Hardy-Weinberg equilibrium were observed, with the exception of DXS101 locus. Allele frequencies and parameters of forensic interest for each X-STR were calculated. High mean exclusion chance and power of discrimination values were obtained by combining these 10 X-linked markers. Population comparisons (exact test of population differentiation; pairwise genetic distances) were carried out and low genetic distances were found between our sample and those from other Spanish or European regions.     

Mitochondrial DNA control region polymorphism in the population of Alagoas state, north-eastern Brazil

The sequences of the two hypervariable (HV) segments of the mitochondrial DNA (mtDNA) control region were determined in 167 randomly selected, unrelated individuals living in the state of Alagoas, north-eastern Brazil. One hundred and forty-five different haplotypes, associated with 139 variable positions, were determined. More than 95% of the mtDNA sequences could be allocated to specific mtDNA haplogroups according to the mutational motifs. Length heteroplasmy in the C-stretch HV1 and HV2 regions was observed in 22 and 11%, respectively, of the population sample. The genetic diversity was estimated to be 0.9975 and the probability of two random individuals presenting identical mtDNA haplotypes was 0.0084. The most frequent haplotype was shared by six individuals. All sequences showed high-quality values and phantom mutations were not detected. The diversity revealed in the mitochondrial control region indicates the importance of this locus for forensic casework and population studies within Alagoas, Brazil.     

Mitochondrial diversity of a northeast German population sample

Mitochondrial DNA sequences of the hypervariable regions HV I and HV II were analyzed in 300 unrelated individuals born and living in the northeast corner of Germany (Western Pomerania) to generate a database for forensic identification purposes in this region. Sequence polymorphism were detected using PCR and direct sequencing analysis. A total of 242 different haplotypes were found as determined by 147 variable positions. The most frequent haplotype (263G, 315.1C) was found in 10 individuals and is also the most common sequence in Europe. Three other haplotypes were shared by 5 individuals, 2 sequences by 4, 8 haplotypes by 3, 15 sequences by 2 persons, and 213 sequences were unique. The genetic diversity was estimated to be 0.99 and the probability of two random individuals showing identical mitochondrial DNA (mtDNA) haplotypes is 0.6%. A comparison with other studies from Germany showed only little differences in the distribution of haplogroups. Nevertheless, one frequent haplotype in northeast Germany (five unrelated individuals) could only rarely be found in other German and European regions. Our results may indicate that despite a high admixture proportion in the German population some regions could demonstrate certain characteristic features.     

Mitochondrial sequence haplotype in the Japanese population

Sequence polymorphysms of the mitochondrial DNA (mtDNA) control region, hypervariable regions I and II, from 50 unrelated Japanese were determined by PCR amplification and cycle sequencing.     

Detection of sequence variation in the HVII region of the human mitochondrial genome in 689 individuals using immobilized sequence-specific oligonucleotide probes

We have developed a rapid, immobilized probe-based assay for the detection of sequence variation in the hypervariable segment II (HVII) of the mitochondrial DNA (mtDNA) control region. Using a panel of 17 sequence-specific oligonucleotide (SSO) probes immobilized on nylon membrane strips, we typed 689 individuals from four population groups. The genetic diversity value for each population was calculated from the frequency data, and the frequencies of distinct "mitotypes" in each group were determined. We performed DNA sequence analysis of 129 samples to characterize the sequences associated with "blanks" (absence of probe signals) and weak probe signals. Out of 689 samples, we observed five heteroplasmic samples (excluding the variable C-stretch beginning at position 303) using the immobilized SSO probe panel. The SSO probe strips were used for the analysis of shed hairs and bloodstains from several criminal cases in Sweden, one of which is described here. We conclude that this mtDNA typing system is useful for human identification and significantly decreases casework turnaround time.     

Stature estimation from radiographically determined long bone length in a Spanish population sample

The estimation of stature from of a variety of bones is an important aspect of forensic work. In order to obtain reliable results, it is important to have comparative data obtained from the same population group as the skeletal remains. However, lack of up to date information on the population groups of Southern Europe makes the estimation of stature from bones in this area subject to possible error. In this study, the stature of 104 healthy adults from Spain was measured, and an anteroposterior teleradiograph of the right lower and the right upper limb of every subject in the study was made in order to measure the lengths of the femur, tibia, fibula, humerus, cubitus and ulna. Pearson's regression formulae were obtained for both limbs. In males, we found the femur to be the most accurate predictor of stature (R = 0.851), whereas in females best results were obtained with the tibia (R = 0.876).     

Mitochondrial DNA Validation in a State Laboratory*,†

Abstract: Because of the inception of the FBI Regional mitochondrial DNA (mtDNA) laboratories, many do not see establishing state/local mtDNA processing laboratories as a priority. Yet there is a long‐term need for mtDNA processing that will exceed the capabilities of the FBI Regional mtDNA laboratories and the few other laboratories that are currently processing mtDNA, and that need can be fulfilled by state/local laboratories. Thus, the DNA Unit of the Delaware Office of the Chief Medical Examiner (OCME‐DNA Unit) completed validation of in‐house mtDNA testing in January 2007. The validation plan for mtDNA processing included the following sections: preliminary research, sensitivity and contamination studies, ExoSAP‐IT^® optimization, BigDye^® optimization, sequencing and 310 optimization, sample preparation and extraction optimization, heteroplasmy, mixtures, and reproducibility. All sections of the validation were successfully completed, and mtDNA processing of skeletal remains, teeth, and hairs, as well as blood and buccal reference samples was adopted by the OCME‐DNA Unit.     

Forearm bones and sexual variation in Turkish population

Forensic anthropologists are aware that there are considerable differences between human populations and therefore develop study models for each skeletal population. The purpose of this study was to analyze forearm bones obtained from forensic settings in Turkey. The sample consists of 42 males and 38 females with an average age of 40 and 36 years, respectively. Numerous measurements were taken from the radius and ulna including lengths (in millimeters), midshaft diameters, and epiphyseal breadths (0.01 mm). Individuals with any anomaly and pathology were not included in the investigation. A stepwise analysis, when applied to individual bones, selected only length and midshaft transverse dimension in the radius and length only in the ulna. When the length was excluded from the statistic, head diameter and distal breadth of the radius and distal minimum head and midshaft anteroposterior diameters of the ulna provided the best predicting functions. Classification results were 92% for the radius and 91% for the ulna. For the incomplete bones, the accuracy rates were about 92% and 83%, respectively. In conclusion, a sex determination was made, in different rates of accuracy, in the human skeleton. Correct assessment can vary among populations. Dimorphism in our region forearm bones is greater than American whites. This supported the hypothesis that human variation is diverse, and population difference should be taken into account when osteometric standards are applied to others. Further studies are needed to understand why the forearm is more dimorphic in Turks.     

Distribution of the minutiae in the fingerprints of a sample of the Spanish population

One of the fundamental aspects of the process of identification through fingerprints is the comparison of the minutiae between the fingermark obtained at the scene of the crime and the suspect's corresponding finger. There is no scientific basis in this process that allows the use of numerical standards, such as those kept in different countries, to obtain the identification. The recent mistakes made in the field of dactyloscopy, together with the growing rigor and scrutiny that forensic evidence undergoes in the legislative and scientific areas, have resulted in the need to reconsider some of the basic principles that support this discipline. A probabilistic estimation of the evidential value is especially necessary; therefore, it is indispensable to know and quantify the variability of the features used in the identification process. The sample studied for this research was obtained from 100 Caucasian men and 100 Caucasian women from the Spanish population, which amounts to a total of 2000 fingerprints. The different types of minutiae were located, identified, and quantified visually on the fingerprint, in four sectors, and inside and outside of a circle, whose radius cut, perpendicularly, fifteen ridges starting from the center cut of the axes that defined the sectors. According to the results obtained in this study, through dactyloscopic identification, the weight of the evidence of a minutia, such as the ridge endings, with frequencies between 55% and 65%, according to the area and gender evaluated, cannot be the same as that of a bifurcation or convergence, with frequencies of 13-18% or those of other minutiae that show frequencies lower than 3%. The significant differences found in the topological distribution of the endings, bifurcations, and convergences show the need to take into account, for its demonstrational value, the finger area in which they are evaluated. The significant association observed between the types of minutiae and the different fingers revealed a greater frequency of endings on the thumb and index fingers, and bifurcations and convergences on the middle, ring, and little fingers.     

Mitochondrial DNA in the Central European population. Human identification with the help of the forensic mt-DNA D-loop-base database

Sequencing of mtDNA is an advanced method for the individualisation of traces. Disadvantages of this method are expensive and time-consuming analysis and evaluation procedures as well as the necessary stock of population-genetic data which is still insufficient. Central European institutes of forensic medicine from Germany, Austria, and Switzerland have been working together since the beginning of 1998 to establish a mtDNA database. The aim is to build up a large stock of forensically established data and provide population-genetic data for frequency investigations, which will serve as a basis for expert opinions and scientific research. Good data quality is ensured by using original sequences only. Ring tests, which have been conducted to enhance analytical reliability, revealed a high correspondence rate of the analytical results obtained by the individual member institutes. Today 1410 sequences are available for comparison, of which 1285 sequences in the HV1 and HV2 regions cover the full ranges from 16051 to 16365 and from 73 to 340 (according to Anderson). The major part is formed by Central European sequences comprising 1256 data sets from Germany, Austria, and Switzerland. Today the database contains sequences from a total of 12 European, six African and three Asian countries including 100 sequences from Japan. This paper is aimed at discussing the individualisation potentials of mtDNA as well as the possibilities and limits of ethnic differentiation by means of pairwise sequence differences on the basis of the data stock available.