The effect of following best practice reporting recommendations on legal and community evaluations of forensic examiners reports.

Commentators have recommended that forensic scientists' reports contain various disclosures to facilitate comprehension. However, little research has explored whether following best practice recommendations for disclosure impacts on receivers' impressions of the evidence. We examined whether forensic science reports that are more compliant with these best practice recommendations reduced overvaluing of the evidence and sensitized legal and community decision-makers to evidence quality. Across three experiments, 240 legal practitioners/trainees and 566 community decision-makers were presented with a fingerprint or footwear report that was either compliant or non-compliant with best practice recommendations. Participants were then asked to make evaluations and decisions based on the report. We found mixed effects of report compliance. Report compliance affected community participant's evaluations of the persuasiveness of the evidence but had limited impact on the judgments of legal practitioners/trainees. When presented with compliant reports, we found that community participants regarded unknown reliability evidence as less reliable and less persuasive than high reliability evidence, suggesting disclosures helped reduce overvaluing of the evidence and create sensitivity to differences in evidence quality. These results suggest compliance with reporting recommendations does affect community impressions, while only minimally influencing legal impressions of forensic science evidence. The costs and/or benefits of this outcome require further examination.

Following the transfer of DNA: How does the presence of background DNA affect the transfer and detection of a target source of DNA?

DNA transfer is of increasing importance in crime scene situations, partly due to analytical techniques detecting profiles in ever declining amounts of DNA. Whereas the focus has previously been DNA transfer of target sources, the effects of background DNA on transfer and detection of DNA after multiple contact situations have been much less investigated. This study measured the transfer and detection rates of a specific DNA source in the presence of background DNA sources. The presence of background DNA influenced the transfer of DNA differently depending on the combination of biological material and surface type. The detection of a profile from the target DNA decreased after multiple contact situations, due to the reduced total and relative quantity of target DNA, and the increasing complexity of the mixture. The results of this study contribute to a greater understanding of the effects of background DNA sources on DNA transfer and detection.

Development of an Italian RM Y-STR haplotype database: Results of the 2013 GEFI collaborative exercise.

Recently introduced rapidly mutating Y-chromosomal short tandem repeat (RM Y-STR) loci, displaying a multiple-fold higher mutation rate relative to any other Y-STRs, including those conventionally used in forensic casework, have been demonstrated to improve the resolution of male lineage differentiation and to allow male relative separation usually impossible with standard Y-STRs. However, large and geographically-detailed frequency haplotype databases are required to estimate the statistical weight of RM Y-STR haplotype matches if observed in forensic casework. With this in mind, the Italian Working Group (GEFI) of the International Society for Forensic Genetics launched a collaborative exercise aimed at generating an Italian quality controlled forensic RM Y-STR haplotype database. Overall 1509 male individuals from 13 regional populations covering northern, central and southern areas of the Italian peninsula plus Sicily were collected, including both "rural" and "urban" samples classified according to population density in the sampling area. A subset of individuals was additionally genotyped for Y-STR loci included in the Yfiler and PowerPlex Y23 (PPY23) systems (75% and 62%, respectively), allowing the comparison of RM and conventional Y-STRs. Considering the whole set of 13 RM Y-STRs, 1501 unique haplotypes were observed among the 1509 sampled Italian men with a haplotype diversity of 0.999996, largely superior to Yfiler and PPY23 with 0.999914 and 0.999950, respectively. AMOVA indicated that 99.996% of the haplotype variation was within populations, confirming that genetic-geographic structure is almost undetected by RM Y-STRs. Haplotype sharing among regional Italian populations was not observed at all with the complete set of 13 RM Y-STRs. Haplotype sharing within Italian populations was very rare (0.27% non-unique haplotypes), and lower in urban (0.22%) than rural (0.29%) areas. Additionally, 422 father-son pairs were investigated, and 20.1% of them could be discriminated by the whole set of 13 RM Y-STRs, which was very close to the theoretically expected estimate of 19.5% given the mutation rates of the markers used. Results obtained from a high-coverage Italian haplotype dataset confirm on the regional scale the exceptional ability of RM Y-STRs to resolve male lineages previously observed globally, and attest the unsurpassed value of RM Y-STRs for male-relative differentiation purposes.

RNA/DNA co-analysis from human menstrual blood and vaginal secretion stains: results of a fourth and fifth collaborative EDNAP exercise.

The European DNA Profiling Group (EDNAP) organized a fourth and fifth collaborative exercise on RNA/DNA co-analysis for body fluid identification and STR profiling. The task was to identify dried menstrual blood and vaginal secretion stains using specific RNA biomarkers, and additionally test 3 housekeeping genes for their suitability as reference genes. Six menstrual blood and six vaginal secretion stains, two dilution series (1/4-1/64 pieces of a menstrual blood/vaginal swab) and, optionally, bona fide or mock casework samples of human or non-human origin were analyzed by 24 participating laboratories, using RNA extraction or RNA/DNA co-extraction methods. Two novel menstrual blood mRNA multiplexes were used: MMP triplex (MMP7, MMP10, MMP11) and MB triplex (MSX1, LEFTY2, SFRP4) in conjunction with a housekeeping gene triplex (B2M, UBC, UCE). Two novel mRNA multiplexes and a HBD1 singleplex were used for the identification of vaginal secretion: Vag triplex (MYOZ1, CYP2B7P1 and MUC4) and a Lactobacillus-specific Lacto triplex (Ljen, Lcris, Lgas). The laboratories used different chemistries and instrumentation and all were able to successfully isolate and detect mRNA in dried stains. The simultaneous extraction of RNA and DNA allowed for positive identification of the tissue/fluid source of origin by mRNA profiling as well as a simultaneous identification of the body fluid donor by STR profiling, also from old and compromised casework samples. The results of this and the previous collaborative RNA exercises support RNA profiling as a reliable body fluid identification method that can easily be combined with current STR typing technology.

Additional Y-STRs in Forensics: Why, Which, and When.

Male-specific DNA profiling using nonrecombining Y-chromosomal genetic markers is becoming ubiquitous in forensic genetics, with many laboratories and jurisdictions taking advantage of the benefits that Y-chromosome short tandem repeat (Y-STR) profiling can bring. The current suite of 9-17 core Y-STRs, available as commercial kits, perform adequately for identifying male lineages in many populations, a feature highly suitable for excluding a male suspect from involvement in crimes such as sexual assaults where autosomal STR profiling is often troubled. However, there is a growing need to achieve higher resolution in paternal-lineage differentiation as adventitious matches between unrelated males are becoming increasingly common with the increasing size of Y-STR haplotype-frequency databases. Furthermore, with the currently used Y-STRs, male relatives (both close and distant) usually cannot be separated, marking a strong limitation in forensic applications as conclusions cannot be drawn on the individual level as desired. Performing Y-chromosome analysis in familial testing, which outperforms autosomal STR profiling in certain deficiency cases, with the current Y-STR sets can be troubled by mutations that complicate relationship-probability estimations. To overcome these limitations, considerable research has been performed over recent years to identify and characterize additional Y-STRs. This review summarizes the forensic performance of current sets of Y-STRs, points out their limitations in the three main areas of forensic Y-STR applications (male-lineage differentiation, male-relative differentiation, and paternity/familial testing), and discusses why and which additional Y-STRs are suitable to improve forensic Y-chromosome analysis in the future.

Locked nucleic acids in PCR primers increase sensitivity and performance.

The incorporation of locked nucleic acids (LNAs) into oligonucleotide primers has been shown to increase template binding strength and specificity for DNA amplification. Real-time PCR and DNA sequencing have been shown to be significantly enhanced by the use of LNAs. Theoretically, increasing primers' binding strength may also increase the sensitivity of conventional PCR, reducing minimum template requirements. We compared LNA-modified PCR primers with their standard DNA counterparts for amplification sensitivity with template amounts as low as 5 pg. Although the results are highly dependent on the design of the LNA primers, large increases in peak height can be achieved from as little as 75 pg, as well as clearer and more complete profiles. Increased amplification success with lower template amounts may also be seen. Additionally, the use of LNAs can enhance multiplexing. Thus, incorporating LNAs into PCR primers can increase amplification success, sensitivity, and performance under a wide range of conditions.