Implementation of a robotized real-time PCR setup for the use of the Quantifiler™ Human DNA Quantification Kit

Human DNA quantification occupies a central role within the DNA analytic process of forensic casework samples as DNA quantification results have an important impact on the quality of the short tandem repeat data. Manual processing for the setup of quantification reactions can be time consuming and labor intensive. Therefore automation of quantitative real-time PCR setup was an important component of our DNA-analysis automation concept. Here we show the implementation of a robotized setup for the Quantifiler™ Human DNA Quantification Kit.     

Automated DNA casework workflow: A retrospective study of the first implementation of FIDL at the Netherlands Forensic Institute

FIDL is a fast and automated DNA identification line which represents a series of software solutions automating the process from raw capillary electrophoresis data to reporting. This retrospective study provides insight in the numbers of cases, turnaround time, results compared to the standard workflow and the benefits automation has in a large volume workflow.     

Diagnostic Value of PSA and AP Tests for the Detection of Spermatozoa in Postmortem Swabs from the Genital and Anal Region in Males

The aim of this study was to clarify whether positive results for prostate‐specific antigen (PSA) and acid phosphatase (AP) occur in postmortem swabs from the genito‐anal region in males (n = 80; 4 regions) and females (n = 20; 3 regions) and to calculate the positive predictive value (PPV) concerning the presence of spermatozoa. In male subjects, the highest incidence of positive test results was found in urethral swabs (PSA 76%, AP 71%) and the lowest frequencies appeared in perianal and rectal swabs (15–20%). Microscopic evaluation for spermatozoa was positive between 39% in urethral swabs and 1% in rectal swabs. PPV regarding positive identification of spermatozoa was 33.3% for PSA and 31.5% for AP. The combination of both tests yielded a PPV of 38.2%. In female cases, no spermatozoa were identified, and one case was PSA‐ and AP‐positive in perianal swabs. Our findings indicate that PSA and AP tests are of limited value for the postmortem detection of spermatozoa in male subjects.     

Could Secondary DNA Transfer Falsely Place Someone at the Scene of a Crime?,

The occurrence of secondary DNA transfer has been previously established. However, the transfer of DNA through an intermediary has not been revisited with more sensitive current technologies implemented to increase the likelihood of obtaining results from low‐template/low‐quality samples. This study evaluated whether this increased sensitivity could lead to the detection of interpretable secondary DNA transfer profiles. After two minutes of hand to hand contact, participants immediately handled assigned knives. Swabbings of the knives with detectable amounts of DNA were amplified with the Identifiler^® Plus Amplification Kit and injected on a 3130xl. DNA typing results indicated that secondary DNA transfer was detected in 85% of the samples. In five samples, the secondary contributor was either the only contributor or the major contributor identified despite never coming into direct contact with the knife. This study demonstrates the risk of assuming that DNA recovered from an object resulted from direct contact.     

A Computational Framework for Age‐at‐Death Estimation from the Skeleton: Surface and Outline Analysis of 3D Laser Scans of the Adult Pubic Symphysis

In forensic anthropology, age‐at‐death estimation typically requires the macroscopic assessment of the skeletal indicator and its association with a phase or score. High subjectivity and error are the recognized disadvantages of this approach, creating a need for alternative tools that enable the objective and mathematically robust assessment of true chronological age. We describe, here, three fully computational, quantitative shape analysis methods and a combinatory approach that make use of three‐dimensional laser scans of the pubic symphysis. We report a novel age‐related shape measure, focusing on the changes observed in the ventral margin curvature, and refine two former methods, whose measures capture the flatness of the symphyseal surface. We show how we can decrease age‐estimation error and improve prior results by combining these outline and surface measures in two multivariate regression models. The presented models produce objective age‐estimates that are comparable to current practices with root‐mean‐square‐errors between 13.7 and 16.5 years.     

法医物证DNA自动化检验技术体系的研究

目的建立自动化工作站同步提取不同种类涉案法医生物检材DNA的新方法。方法选用TECAN Freedom EVO100．4、75—2型自动化提取、加样工作站,采用磁珠法及Chelex-100法对各类涉案生物检材进行DNA提取、PCR扩增、毛细管电泳检测其STR分型,进行比较测试。在“全国公安机关DNA数据库应用系统”中建立并应用实验室信息管理系统（LIMS）模拟实施规范化DNA检案。结果1552份各类检材,采用工作站-磁珠法提取DNA效果最佳,STR检测成功率为95％,工作站-Chelex法为88％;二者分别与其手工提取法比较,成功率无明显差异。92个样本同期检测,自动化工作站较手工操作DNA检案时间可缩减1．25倍。结论工作站域珠法提取涉案检材DNA,可获得满意的STR分型结果。应用LIMS管控,可有效防控污染,明显提高检案效率及鉴定质量。     

An investigation into the relationship between edge counts and the variability of the refractive index of glass. Part I: Edge morphology

The effect of glass fragment morphology on the variability of refractive index values in five different glass objects was investigated. Data are presented that suggest that the variability of refractive index values is increased when fragment edge morphology becomes unsuitable for phase contrast microscopy.     

Validation and casework application of a Y chromosome specific STR multiplex

A series of validation experiments was performed for a Y chromosome specific STR multiplex system following the suggestions made by the Technical Working Group DNA Analysis Methods (TWGDAM). The multiplex PCR products were detected on Perkin-Elmer 373 and 377 automated sequencers using two labeling colors. No problems regarding the stability, robustness and sensitivity of the Y STR multiplex were observed. Mixture studies revealed a cut off rate similar to autosomal STRs for mixtures of male DNAs and no interference of any female admixture. The comparison of the Y STR results to the autosomal typing results for 56 nonprobative semen stains and swabs, showed a slightly higher success rate in detecting the semen donor’s alleles for the Y STR multiplex. Two examples are shown to illustrate the usefulness of Y STR typing for DNA mixtures. In one case the Y STR results confirmed an isolated exclusion; in the other case, the interpretation of a mixture was clarified since the Y STR results proved the presence of DNA from at least two semen donors. Y STR typing is a valuable addition to the forensic DNA testing panel.     

Applicability of DNA analysis on adhesive tape in forensic casework

Adhesive tape is commonly used in crimes and is often the subject of forensic evaluation. DNA analysis of adhesive tape can provide DNA profiles of suspects. The object of this study was to evaluate the applicability of DNA analysis on adhesive tape samples in forensic casework. We retrospectively reviewed all cases involving adhesive tape or similar items received by our institute for DNA analysis during the past 11 years. From 100 forensic cases reviewed, 150 adhesive tape samples were examined. A total of 98 DNA profiles were obtained from these samples. Sixty-two of the profiles provided feasible case-relevant information. In conclusion, DNA profiling of adhesive tape samples can be useful in a variety of forensic cases.