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1.
Species identification is fundamental to wildlife forensic practice. The desirability of molecular genetic methods is increasing rapidly. The sequence of a marker, rather than its particular diagnostic nucleotides, provides greater safety through comparisons between intra- and inter-specific pairwise genetic distances. However, it has not been well described how reliability of species assignment is influenced by distance computing methods and reference sample sizes. In this study, the influences were tested using 12 species from 4 genera of passerine birds and the sequences of partial Cytochrome b (Cyt b) and Cytochrome Oxidase subunit I (COI) genes. Results showed that different substitution types have different outcomes of pairwise genetic distance estimation and this influences the risk of false inclusion and exclusion. Transition (Ts) is the most effective substitution type to reveal optimal species resolution for both Cyt b and COI gene fragments no matter whether K2P and p-distance are used. Sample size required to accurately estimate pairwise distance is essentially determined by the genetic diversity of a species in reference to a given strictness of predefined acceptable accuracy. These findings suggest that for future forensic work on birds by use of Cyt b and COI gene fragments, transition should be used exclusively for marker validation and identification practice when targeting closely related species. Meanwhile, the reference database should sufficiently represent overall genetic diversity of the species. The minimum sample size should be estimated based on existing knowledge of genetic diversity. Special caution should be used for species assignment when only several reference data are available for animals that are considered likely to have high genetic diversity.  相似文献   

2.
The utility of the forensically important Sarcophagidae (Diptera) for time since death estimates has been severely limited, as morphological identification is difficult and thermobiological histories are inadequately documented. A molecular identification method involving the sequencing of a 658-bp ‘barcode’ fragment of the mitochondrial cytochrome oxidase subunit I (COI) gene from 85 specimens, representing 16 Australian species from varying populations, was evaluated. Nucleotide sequence divergences were calculated using the Kimura-two-parameter distance model and a neighbour-joining phylogenetic tree generated. All species were resolved as reciprocally monophyletic, except Sarcophaga dux. Intraspecific and interspecific variation ranged from 0.000% to 1.499% (SE = 0.044%) and 6.658% to 8.983% (SE = 0.653%), respectively. The COI ‘barcode’ sequence was found to be suitable for the molecular identification of the studied Australian Sarcophagidae: 96.5% of the examined specimens were assigned to the correct species. Given that the sarcophagid fauna is poorly described, it is feasible that the few incorrectly assigned specimens represent cryptic species. The results of this research will be instrumental for implementation of the Australian Sarcophagidae in forensic entomology.  相似文献   

3.
Species identification techniques commonly utilized in Australian Forensic Science laboratories are gel immunodifussion antigen antibody reactions and hair comparison analysis. Both of these techniques have significant limitations and should be considered indicative opinion based tests. The Barcode of Life Initiative aims to sequence a section of DNA (~648 base pairs) for the Cytochrome Oxidase I mitochondrial gene (COI) in all living species on Earth, with the data generated being uploaded to the Barcode of Life Database (BOLD) which can then be used for species identification. The COI gene therefore offers forensics scientists an opportunity to use the marker to analyze unknown samples and compare sequences generated in BOLD. Once sequences from enough species are on the database, it is anticipated that routine identification of an unknown species may be possible. However, most forensic laboratories are not yet suited to this type of analysis and do not have the expertise to fully interpret the implications of matches and non matches involving a poorly sampled taxa (for example where there are cryptic species) and in providing the required opinion evidence. Currently, the use of BOLD is limited by the number of relevant species held in the database and the quality assurance and regulation of sequences that are there. In this paper, the COI methodology and BOLD are tested on a selection of introduced and Australian mammals in a forensic environment as the first step necessary in the implementation of this approach in the Australian context. Our data indicates that the COI methodology performs well on distinct species but needs further exploration when identifying more closely related species. It is evident from our study that changes will be required to implement DNA based wildlife forensics using the BOLD approach for forensic applications and recommendations are made for the future adoption of this technology into forensic laboratories.  相似文献   

4.
Forensic scientists are increasingly using DNA to identify the species of a tissue sample. However, little attention has been paid to basic experimental design issues such as replication and the selection of taxa when designing a species diagnostic test. We present an example using the forensically important fly genus Lucilia in which an increasingly larger sample size revealed that species diagnosis based on the commonly used cytochrome oxidase I gene (COI) was less straightforward than we initially thought. This locus may still be useful for diagnosing Lucilia specimens, but additional knowledge other than the genotype will be required to reduce the list of candidate species to include only forms that can be distinguished by COI. We believe that these results illustrate the importance of study design and biological knowledge of the study species when proposing a DNA-based identification test for any taxonomic group. Electronic supplementary material Supplementary material is available for this article at and is accessible to authorized users.  相似文献   

5.
Molecular biological species identification of forensically important flies distributed in Iwate prefecture, Japan, was carried out, and the utility of this method was evaluated. The dipteran nymphs that were early colonizers on the corpse were reared to adult and morphologically identified. Meanwhile they were sequenced over a 304 base pair region of the mitochondrial cytochrome oxidase gene subunit I (COI). Six species belonging three genera of Calliphoridae (Calliphora lata, C. vicina, Lucilia cuprina, L. illustris, L. sericata, Chrysomya pinguis) and two species of Sarcophagidae (Parasarcophaga crassipalpis, P. similis) were collected and identified. Each fly had somewhat different ecological features such as seasonal dominance and habitat. The COI sequences of each species were unique and distinguishable from each other, although they showed high homology. Species identification from immature diptera by the DNA sequences was simple and time-saving because there was no need to wait for adult emergence or knowledge of morphological keys, and it provided information about not only postmortem interval but also seasonal and environmental conditions surrounding the corpse.  相似文献   

6.
Screening methods sensitive to movement strategies that increase anterior cruciate ligament (ACL) loads are likely to be effective in identifying athletes at‐risk of ACL injury. Current ACL injury risk screening methods are yet to be evaluated for their ability to identify athletes' who exhibit high‐risk lower limb mechanics during sport‐specific maneuvers associated with ACL injury occurrences. The purpose of this study was to examine the efficacy of two ACL injury risk screening methods in identifying high‐risk lower limb mechanics during a sport‐specific landing task. Thirty‐two female athletes were screened using the Landing Error Scoring System (LESS) and Tuck Jump Assessment. Participants' also completed a sport‐specific landing task, during which three‐dimensional kinematic and kinetic data were collected. One‐dimensional statistical parametric mapping was used to examine the relationships between screening method scores, and the three‐dimensional hip and knee joint rotation and moment data from the sport‐specific landing. Higher LESS scores were associated with reduced knee flexion from 30 to 57 ms after initial contact (P = 0.003) during the sport‐specific landing; however, no additional relationships were found. These findings suggest the LESS and Tuck Jump Assessment may have minimal applicability in identifying athletes' who exhibit high‐risk landing postures in the sport‐specific task examined.  相似文献   

7.
Forensic botany can provide significant supporting evidence during criminal investigations. However, it is still an underutilized field of investigation with its most common application limited to identifying specific as well as suspected illegal plants. The ubiquitous presence of plant species can be useful in forensics, but the absence of an accurate identification system remains the major obstacle to the present inability to routinely and correctly identify trace botanical evidence. Many plant materials cannot be identified and differentiated to the species level by traditional morphological characteristics when botanical specimens are degraded and lack physical features. By taking advantage of a universal barcode system, DNA sequencing, and other biomolecular techniques used routinely in forensic investigations, two chloroplast DNA regions were evaluated for their use as “barcoding” markers for plant identification in the field of forensics. We therefore investigated the forensic use of two non-coding plastid regions, psbA-trnH and trnL-trnF, to create a multimarker system for species identification that could be useful throughout the plant kingdom. The sequences from 63 plants belonging to our local flora were submitted and registered on the GenBank database. Sequence comparison to set up the level of identification (species, genus, or family) through Blast algorithms allowed us to assess the suitability of this method. The results confirmed the effectiveness of our botanic universal multimarker assay in forensic investigations.  相似文献   

8.

We evaluated whether MinION, an inexpensive portable sequencer, can be used to identify the origin of crude drugs derived from animals. Standard and nonstandard crude drugs with different species of origin were examined. In addition, standards mixed with nonstandard samples were used. As a target gene, cytochrome c oxidase I was amplified and sequenced. The fast mode results had a slightly lower match ratio than high-accuracy mode, but the animals of origin were correctly determined by BLAST for all samples. For antler velvet derived from Rangifer tarandus, even when the sequences were aligned based on Cervus elaphus, the animal of origin was determined correctly. Minor contents could be detected from mixtures of two animals, if the mixtures contained at least 19:1 mtDNA when the coverage allele-fraction threshold was 0.05. By contrast, in fast mode, two sequences could not be separated due to the low accuracy of the base-calling for each read. For fieldwork, the species of origin of crude drugs could be identified with only simple DNA extraction and library preparation. Therefore, MinION appears to be a convenient tool for identifying the origins of crude drugs derived from animals.

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9.
13种虫媒病毒基因芯片检测方法的建立   总被引:3,自引:1,他引:2  
目的 建立能对虫媒病毒进行属水平筛查及种水平鉴定的基因芯片技术,为虫媒病毒提供一种高通量的检测与鉴定技术.方法 从GenBank获得13种虫媒病毒的全部基因组序列,利用Clustal X和BLAST等比对软件,设计针对这些虫媒病毒所在的3个属的通用寡核苷酸探针,用于病毒的属水平筛查.然后设计针对每种病毒的特异性寡核苷酸探针,用于每种病毒的检测鉴定.寡核苷酸探针长度均为70mer,将设计好的探针与GenBank上所有病毒基因组序列进行比对,验证其种属特异性,然后进行探针合成和基因芯片制备.所有病毒在BSL-3和BSL-2实验室进行培养,待检病毒核酸通过锚定随机PCR进行扩增,经荧光染料标记后与基因芯片杂交,利用激光共聚焦扫描仪扫描后进行结果分析.结果 所检测的13种虫媒病毒均可获得种特异性及所在属的属特异性杂交信号,非特异性杂交信号不明显.分别用两种不同的病毒混合后,进行模拟混合病毒感染,也得到了相应的特异杂交信号.结论 初步建立了13种虫媒病毒的基因芯片检测技术,可用于这些病毒的属水平筛查及种水平鉴定,并且可以进行混合感染标本的检测,为虫媒病毒的检测与鉴定提供了一种新的手段.  相似文献   

10.
In forensic cases, blood alcohol is routinely requested. Although the standard method for alcohol analysis is the gas chromatographic technique, the instrument is not available in some hospital laboratories. As a result, there is an attempt to use an automatic alcohol analyzer for preliminary testing. However, a barrier to application is the putrefactive changes of post-mortem specimens that might interfere with the analysis and cause false positive results. This study aims to determine the accuracy of an automatic VITROS alcohol analysis for post-mortem specimens. Blood and urine samples were modified to putrefactive appearances and analyzed for ethanol by VITROS. The measured amounts were compared with the added ethanol amounts. The results indicated that samples with fresh hemolysis and putrefactive volatile substances gave false high ethanol results. Plasma with a high glucose content in the presence of Escherichia coli or Candida albicans did not show neo-ethanol formation. Indeed, all samples left at room temperature prior to analysis showed false low detected ethanol amounts. Thus, VITROS alcohol analysis has limitations leading to inaccurate post-mortem specimen analysis. In addition, loss of alcohol from corpses and specimen containers also has an impact on the accuracy of alcohol results in post-mortem cases.  相似文献   

11.
The population origin of an individual is often requested to be determined from specimens left at a crime scene for identifying a suspect and individual identity. The melanocortin 1 receptor gene (MC1R) and P gene are associated with human pigmentation. Although several studies have reported that these genes are highly polymorphic in human populations, it is unclear if the allele variants can be used to determine the population origin of an individual. We aimed to determine the ethnic origin of an individual by using single nucleotide polymorphisms (SNPs). Eighteen SNPs in the MC1R gene and P genes were genotyped in 52 individuals by the direct sequencing method, and 4 SNPs (MC1R gene: R163Q and P gene: IVS5 + 1001, IVS13 + 113, and H615R) were selected on the basis of differences in frequencies. Subsequently, we genotyped these four SNPs in 422 volunteers from six ethnically defined populations using a polymerase chain reaction-based assay. The results revealed that the allele variants were present with high frequencies in Asian populations but were low in European and African populations. On the basis of these results, we defined a specific combination of a genotype (R163Q) and a diplotype group (IVS5 + 1001, IVS13 + 113, and H615R). This study indicates that the specific combination of a genotype and a diplotype group would be effective in estimating the population origin of an individual from a list of population groups. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.  相似文献   

12.
The goals of this study were to determine the prevalence and determinants of false‐positive exercise tests in athletes. Data from all athletes who visited the Department of Sport Medicine for assessment of sports eligibility during a 1.5‐year period were reviewed retrospectively. Potential determinants of (false) positive test results that were evaluated included demographics, cardiovascular risk factors, sports characteristics, resting electrocardiogram (ECG) abnormalities, and exercise capacity. Data from 1298 athletes were included. In 53 athletes (4.1%), the exercise ECG was classified as positive. Among 38 athletes who were referred to a sports cardiologist for further diagnostic evaluation, 36 (95%) were classified as having a false‐positive test result and 2 athletes (5%) required coronary revascularization. Athletes with a false‐positive test were older than athletes with a negative test (53 ± 8 vs 45 ± 13 years, P = 0.03). In conclusion, exercise electrocardiography has a low positive predictive value in asymptomatic recreational and competitive athletes, with a false‐positive test result being associated with higher age. Given the relatively high prevalence of false‐positive test results in this population, efforts should be made to develop strategies aimed at identifying false‐positive test results in a simple noninvasive manner.  相似文献   

13.
Extemporary (EXT) analysis is unavoidable in establishing the tumor diagnosis, operability and the extent of the operation. Alternative approach is cytologic analisis which, because of its simple methodology, provides results even faster. In this paper, the results of cytologic imprints (CI) and EXT finding were compared with definite histopathologic diagnosis (HDP) to determine the value of both methods. A total of 109 samples obtained during 55 thoracotomies were analyzed. Eighty eight specimens were analyzed simultaneously by CI Method and in frozen sections. Twenty-one sample was analyzed only by cytologic methods and the results of standard CI were compared with definite HDP. After being processed for EXT diagnosis, intraoperative specimens were imprinted on glass slides, air-dried and stained by May-Grünwald--Giemsa Method. In cytologic analysis there were no false negative results, but there were 7 false positives. The overall diagnostic accuracy was 93.6%, sensitivity and negative predictive value was 100%, specificity was 91.1% and positive predictive value was 81.8%. Diagnostic accuracy of frozen sections was 98.8% also without false negatives and with one false positive finding with sensitivity and negative predictive value of 100%, specificity of 98.4% and positive predictive value of 95%. These results corresponded to the results of other studies and confirmed the efficacy of CI method, which could be used either simultaneously with EXT diagnosis as a complementary or as an alternative method in the hospitals where EXT analysis is not used. However, imprint cytology demands an experienced cytologist and could be used only in hospitals with well organized cytologic service.  相似文献   

14.

Ethyl glucuronide (EtG) is a direct alcohol marker, which is widely used for clinical and forensic applications, mainly for abstinence control. However, the instability of EtG in urine against bacterial degradation or the post-collectional synthesis of EtG in contaminated samples may cause false interpretation of EtG results in urine samples. This study evaluates the potential of sodium azide in tubes used for urine collection to hinder degradation of ethyl glucuronide by bacterial metabolism taking place during growth of bacterial colonies. The tubes are part of a commercial oral fluid collection device. The sampling system was tested with different gram-positive and gram-negative bacterial species previously observed in urinary tract infections, such as Escherichia coli, Staphylococcus aureus, Enterecoccus faecalis, Staphylococcus epidermidis, Klebsiella pneumoniae, Enterobacter cloacae, and Pseudomonas aeruginosa. Inhibition of bacterial growth by sodium azide, resulting in lower numbers of colony forming units compared to control samples, was observed for all tested bacterial species. To test the prevention of EtG degradation by the predominant pathogen in urinary tract infection, sterile-filtered urine and deficient medium were spiked with EtG, and inoculated with E. coli prior to incubation for 4 days at 37 °C in tubes with and without sodium azide. Samples were collected every 24 hours, during four consecutive days, whereby the colony forming units (CFU) were counted on Columbia blood agar plates, and EtG was analyzed by LC-MS/MS. As expected, EtG degradation was observed when standard polypropylene tubes were used for the storage of contaminated samples. However, urine specimens collected in sodium azide tubes showed no or very limited bacterial growth and no EtG degradation. As a conclusion, sodium azide is useful to reduce bacterial growth of gram-negative and gram-positive bacteria. It inhibits the degradation of EtG by E. coli and can be used for the stabilization of EtG in urine samples.

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15.
Objective. To assess hyaline cartilage of cadaveric ankles using different magnetic resonance (MR) imaging techniques and various methods of measurement. Design and patients. Cartilage thicknesses of the talus and tibia were measured in ten cadaveric ankles by naked eye and by digitized image analysis from MR images of fat-suppressed T1-weighted gradient recalled (FS-SPGR), sequences and pulsed transfer saturation sequences with (FS-STS) and without fat-suppression (STS); these measurements were compared with those derived from direct inspection of cadaveric sections. The accuracy and precision errors were evaluated statistically for each imaging technique as well as measuring method. Contrast-to-noise ratios of cartilage versus joint fluid and marrow were compared for each of the imaging sequences. Results. Statistically, measurements from FS-SPGR images were associated with the smallest estimation error. Precision error of measurements derived from digitized image analysis was found to be smaller than that derived from naked eye measurements. Cartilage thickness measurements in images from STS and FS-STS sequences revealed larger errors in both accuracy and precision. Interobserver variance was larger in naked eye assessment of the cartilage. Contrast-to-noise ratio of cartilage versus joint fluid and marrow was higher with FS-SPGR than with FS-STS or STS sequences. Conclusion. Of the sequences and measurement techniques studied, the FS-SPGR sequence combined with the use of digitized image analysis provides the most accurate method for the assessment of ankle hyaline cartilage.  相似文献   

16.
Chew FS  Kline MJ 《Radiology》2001,218(1):211-214
PURPOSE: To evaluate the diagnostic yield of computed tomography (CT)-guided percutaneous needle aspiration procedures in the setting of suspected spontaneous infectious diskitis and to assess the usefulness of concurrent cytologic examination as a supplement to microbiologic evaluation. MATERIALS AND METHODS: A retrospective study was performed to evaluate 105 consecutive CT-guided percutaneous disk space aspiration procedures in 92 patients suspected of having spontaneous (non-postoperative) infectious diskitis. Our criterion standard for the presence of active infection was the identification of a pathogen either from the CT-guided aspiration specimen or from a surgical specimen. All cases had microbiologic analysis, 78 cases had cytopathologic analysis, and 31 cases had open surgery. RESULTS: Microbiologic analysis of the CT-guided percutaneous aspiration specimens was positive in 39 of 43 cases proved to have active infections, with four false-negative and no false-positive cases (sensitivity, 91%; specificity, 100%). The false-negative cases were all active fungal infections identified from surgical specimens. Adding cytopathologic analysis to microbiologic analysis improved sensitivity but reduced specificity. The most common pathogens were species of Staphylococcus, Streptococcus, Candida, and Mycobacterium. All 30 active bacterial infections were identified with the CT-guided procedures, but only five of nine fungal infections were identified. CONCLUSION: CT-guided percutaneous needle aspiration is an accurate method for identifying active bacterial disk space infections but is less reliable for identifying fungal infections.  相似文献   

17.
Objective. To identify an optimal pulse sequence for in vitro imaging of hyaline cartilage at 0.5 T. Materials and methods. Twelve holes of varying diameter and depth were drilled in cartilage of two pig knees. These were submerged in saline and scanned with a 0.5-T MR system. Sixteen T1-weighted gradient echo (GE), two T2-weighted GE, and 16 fast spin echo sequences were used, by varying repetition time (TR), echo time (TE), flip angle (FA), echo train length, profile order, and by use of fat saturation. Contrast-to-noise ratios (CNR) of cartilage versus saline solution and cartilage versus subchondral bone were measured. Cartilaginous lesions were evaluated separately by three independent observers. Interobserver variability and correlation between the quantitative and qualitative analyses were calculated. Results. The mean CNRs of two specimens of cartilage versus saline solution ranged from 6.3 (±2.1) to 27.7 (±2.5), and those of cartilage versus subchondral bone from 0.3 (±0.2) to 22.5 (±1.4). The highest CNR was obtained with a T1-weighted spoiled 3D-GE technique (TR 65 ms, TE 11.5 ms, FA 45°). The number of lesions observed per sequence varied from 35 to 69. Observer agreement was fair to good. The T1-weighted spoiled GE sequences with a TR of 65 ms, TE of 11.5 ms and FA of 30° and 45° were significantly superior to the other 34 sequences in the qualitative analysis. Conclusion. T1-weighted spoiled 3D-GE sequences with a TR of 65 ms, a TE of 11.5 ms, and a FA of 30–45° were found to be optimal for in vitro imaging of cartilage at 0.5 T.  相似文献   

18.
Proteomic genotyping detects single amino acid polymorphisms to infer the genotype of corresponding non-synonymous SNPs. Like any DNA genotype, these inferences can be used to estimate random match probability. Fingermarks are a common source of biological evidence that is sample limited and a highly variable source of identifying DNA. Genetically variant peptides from fingermarks, that contain single amino acid polymorphisms, are an additional source of identifying genetic information. To discover these peptide biomarkers epidermal corneocytes from 9 subjects were isolated, processed, digested with trypsin and applied to mass spectrometry. The resulting proteomic and matching exome datasets were used to discover, characterize and validate 60 genetically variant peptides. An average of 28.8 ± 4.4 genetically variant peptides were detected from each subject resulting in a total of 264 SNP allele inferences with 260 true and 4 false positives, a false discovery rate of 1.5%. Random match probabilities were estimated using the genotype frequencies from the matching major populations in the 1000 Genomes Project. Estimates ranged up to a value of 1 in 1.7 × 108, with a median probability of 1 in 2.4 × 106. Furthermore, the proteomically-inferred genotypes are likely to be compatible with the STR-based random match probability estimates since the closest STR locus was 2.2 Mb from the nearest GVP-inferred SNP. This project represents a novel mode of genetic information that can be obtained from fingermarks and has the potential to complement other methods of human identification including analysis of ridge patterns or touch DNA.  相似文献   

19.
Autopsies of exhumed cadavers can reveal important evidence for clarification of medical insurance and social issues. This study concerns insects sampled on 22 exhumed cadavers in the Lille area. For each corpse, the species and the stages of development were noted, as well as the time elapsed after burial, the location of the cemetery, the stage of decay and possible preservation treatment. A total of eight Diptera and two Coleoptera species were sampled on the corpses. The relationships between entomofauna and conditions of burial are discussed. Three species were regularly found because of their preference for underground environments or closed environments: Conicera tibialis, typically associated with buried bodies, Leptocera caenosa which is known to be associated with human faeces, water closets, caves and cracked soil pipes, and Ophyra capensis, sometimes found on human bodies kept indoors for several months, where blowflies have not had access. Triphleba hyalinata, which is associated with human bodies in wooden coffins, was found only twice.  相似文献   

20.
目的评价达亚美Techno TwinStation全自动血型仪在交叉配血中的准确性。方法用达亚美Techno TwinStation全自动血型仪对4 500例患者样本进行交叉配血,并与传统的抗人球蛋白试管法、聚凝胺法进行对照试验。结果 (1)200份抗体筛查阴性的标本,三种方法交叉配血试验结果完全相符;4 280份抗体筛查阴性的标本,全自动血型仪和聚凝胺法交叉配血试验结果完全相符;(2)20份抗体筛查阳性标本,交叉配血试验结果为:全自动血型仪法20例不相容,聚凝胺法16例不相容,抗人球蛋白试管法18例不相容,结果比较无统计学差异(P>0.05)。结论达亚美Techno TwinStation血型仪用于交叉配血检测,结果可靠,实验操作规范化、标准化,降低了人为错误的发生率;实验结果可永久保存,便于查询和医疗举证。  相似文献   

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