Statistical Modeling of the Case Information From the Ohio Attorney General's Sexual Assault Kit Testing Initiative

The Ohio Attorney General's Sexual Assault Kit (SAK) Testing Initiative has resulted in nearly 14,000 kits being processed since the initiation of the project in 2012. A logistic regression model was fit to the data from 2500 SAKs in order to determine the probability of obtaining at least one Combined DNA Index System (CODIS) eligible DNA profile based on a number of predictor variables. The probability of obtaining at least one CODIS eligible DNA profile from an SAK varied as a function of (i) days to kit collection following a sexual assault; (ii) years to kit submission to the laboratory for testing following kit collection; (iii) the age of the victim; and (iv) the occurrence of victim‐reported consensual sex around the time of the assault and/or kit collection. These findings demonstrate the utility of the statistical modeling of data obtained from the “forklift” testing approach of sexual assault kits.     

A State Census of Unsubmitted Sexual Assault Kits: Comparing Forensic DNA Testing Outcomes by Geographic and Population Density Characteristics*

A growing number of U.S. cities and states have large numbers of unsubmitted sexual assault kits (SAKs) in police property facilities. Prior research conducted in large urban cities has found that testing these kits yields a sizable number of DNA profiles that meet FBI eligibility for upload to the national criminal DNA database CODIS (Combined DNA Index System) and uploaded profiles return a substantial number of matches to existing criminal profiles in CODIS. It is unknown whether these findings are unique to large urban cities with high crime rates. The purpose of current study was to document forensic testing outcomes from a state census of previously unsubmitted SAKs, which included large urban–suburban centers, as well as smaller cities and rural counties. We inventoried all previously unsubmitted SAKs in Michigan (N = 3422 SAKs) and submitted all kits for forensic DNA testing. A total of n = 1239 SAKs had a DNA profile that met eligibility for upload into CODIS (36.2% unconditional, 56.5% conditional CODIS eligible rate) and n = 585 SAKs yielded a CODIS Hit (17.1% unconditional, 47.2% conditional CODIS hit rate). These rates are consistent with studies from urban areas suggesting approximately half of SAKs tested yield a CODIS profile and approximately half of those uploaded profiles yield a hit. We compared SAK forensic testing outcomes by geographic and population density characteristics, and although rates were often higher in larger metropolitan areas, the obtained rates in micropolitan and rural areas suggest testing is warranted in smaller jurisdictions as well.     

The problem of untested sexual assault kits: why are some kits never submitted to a crime laboratory?

Victims of sexual assault are often advised to seek postassault medical care to have a forensic exam, which includes evidence collection (termed a sexual assault kit [SAK]). After the exam, law enforcement personnel are supposed to submit the SAK to a crime laboratory for analysis. However, recent media reports suggest that in many communities throughout the United States, thousands of SAKs are left untested. Few studies have examined the rate at which law enforcement submits SAKs to crime labs or the factors that may predict them to do so. Thus, the purpose of this exploratory study is twofold: (a) to examine the percentage of SAKs law enforcement submits to crime labs in cases in which a sexual assault nurse examiner (SANE) performed the exam with adult victims and (b) to explore whether assault and law enforcement characteristics predict whether SAKs are submitted to a crime lab. This study found that only 58.6% of the SAKs were submitted to the crime lab within a large Midwestern county. Using binary logistic regression, this study found that kits were significantly as likely to be submitted when there were documented physical (nonanogenital) injuries compared with kits that did not have documented physical injuries. In addition, kits that were handled by a law enforcement agency that had a high level of engagement with the SANE program were significantly as likely to be submitted as law enforcement agencies with a low or medium level of engagement. Kits were significantly less likely to be submitted when victims cleaned themselves after the sexual assault (e.g., bathing). No association was found between kit submission and the victim-offender relationship, suspected drug-facilitated sexual assault, anogenital injury, and when the victim consumed alcohol or drugs before the assault. This article concludes with a discussion of the implications for research and practice.     

Investigation into "normal" background DNA on adult necks: implications for DNA profiling of manual strangulation victims

Others have investigated the role that DNA profiling could play as a method for identifying the perpetrator of manual strangulation. These studies have demonstrated that it is possible to collect offender DNA from the skin surface of a victim following physical contact. It is not known whether nonself biological material is normally present on the skin surface due to adventitious transfer occurring during innocent everyday interactions. To test the hypothesis that detectable amounts of nonself DNA are normally present on the skin surface of healthy adult individuals due to the adventitious transfer of DNA occurring during normal day-to-day social interactions, we designed an experiment in three phases. Phase 1 was used to deduce which DNA collection, extraction, and amplification methods were suited to investigating this question. During phase 2, the neck surface of 24 healthy adult volunteers was swabbed. DNA was extracted using the QIAamp DNA mini kit and amplified using the SGM Plus PCR amplification kit, using 28 PCR cycles. The work carried out during phase 3 involved a simulated assault to investigate primary and secondary transfer of DNA during physical contact. It was found that 23% of neck areas swabbed during phase 2 of this investigation showed nondonor alleles in the resulting DNA profile, with 5% of areas showing six or more nondonor alleles. The results of phase 3 showed that primary, secondary, and zero transfer of victim and/or offender DNA could be observed after physical contact and that alleles from an unknown source could still be detected in this more controlled experiment. The data presented in this paper demonstrate that DNA profiles generated after swabbing the skin surface of healthy adults can include components of an unknown source, present due to adventitious transfer. These components, if present in large quantities, have the potential to interfere with DNA profile interpretation of swabs taken for the investigation of physical assault by DNA profiling.     

Analysis of non-suspect samples lacking visually identifiable sperm using a Y-STR 10-plex

Y-STRs are valuable in the investigation of sexual assaults in which autosomal STR genotype interpretation is challenging. To detect male DNA from compromised sexual assault evidence, 45 non-suspect samples were differentially extracted and analyzed with 10 Y-STRs. These samples were positive for the presence of human seminal fluid, but were negative for spermatozoa by microscopic examination. Y-STR data were obtained in approximately 86.2% of the epithelial or sperm fractions. On samples yielding incomplete profiles, results were obtained on an average of 5 loci per sample. The inability to obtain results may be due to insufficient amplifiable male DNA, PCR inhibition, or unfounded accusations of sexual assault. This study indicates that it is possible to obtain a male STR profile even in the absence of visually identifiable spermatozoa. Furthermore, Y-STR loci should become components of CODIS if they are to be used in solving non-suspect sexual assaults.     

Qiagen's Investigator™ Quantiplex Kit as a Predictor of STR Amplification Success from Low‐Yield DNA Samples,

Qiagen's Investigator? Quantiplex kit, a total human DNA quantitation kit, has a 200‐base pair internal control, fast cycling time, and scorpion molecules containing a covalently linked primer, probe, fluorophore, and quencher. The Investigator? Quantiplex kit was evaluated to investigate a value under which complete short tandem repeat (STR) failure was consistently obtained. Buccal swabs were extracted using the Qiagen QIAamp^® DNA Blood Mini Kit, quantified with the Investigator? Quantiplex kit using a tested half‐volume reaction, amplified with the ABI AmpFlSTR^® Identifiler kit, separated on the 3100Avant Genetic Analyzer, and data analyzed with GeneMapper^® ID v.3.2. While undetected samples were unlikely to produce sufficient data for statistical calculations or CODIS upload (2.00 alleles and 0.82 complete loci on average), data may be useful for exclusionary purposes. Thus, the Investigator? Quantiplex kit may be useful for predicting STR success. These findings are comparable with previously reported data from the Quantifiler? Human kit.     

Biological and DNA evidence in 1000 sexual assault cases

Using a Filemaker-based database (DNA Pro-FILES, Synchrone Infosystème Inc.), we have conducted a large-scale study on 1000 sexual assault (SA) cases where a standardized kit was submitted to our laboratory alone or with other types of exhibits. We looked at the likelihood of obtaining good quality DNA evidence, allegedly from the assailant, according to a number of parameters.The overall proportion of SA cases with DNA evidence is nearly 50%. A little more than 30% of SA kits provided DNA evidence while for 16% of cases DNA evidence could be obtained only from other exhibits.The likelihood of obtaining DNA evidence is approximately 50% in teenager and adult SA cases, but much lower for children 10 years old or younger (15%). In children cases, profiles were found mostly on clothing or skin swabs.The likelihood of obtaining DNA evidence from vaginal swabs remains good for up to 3 days after the assault (from 35% on the first day to 23% on the third day). A DNA profile was obtained from approximately 22% of anal/rectal swabs and 41% of skin swabs taken less than 1 day after the assault. Less than 10% of oral washes provided DNA evidence, all having been collected within 24 h of the assault.We found that in bodily samples, a negative result for acid phosphate (AP) is a poor predictor of the likelihood of obtaining good quality DNA evidence. Approximately 15% of vaginal swabs and 8% of anal swabs negative for AP nevertheless provided good quality DNA evidence.     

Sexual assault cases related to unknown perpetrator: Almost 50% of the analyzed cases corresponded to serial offenders

As a collaborative effort in solving sexual assault cases where there is no suspect, the DNA profiles associated with 273 cases were analyzed. Such cases were submitted from different Criminal Investigation Units belonging to eight judicial departments of Buenos Aires Province. A single NN male profile was recovered from the evidences by differential lysis with DNA IQ System (Promega) and typing with IdentiFiler kit. Comparative analysis of the compiled DNA profiles showed that in 45% of the cases the evidence DNA profile matched in at least two unrelated cases. Associations between groups of unsolved cases provided a valuable tool in aiding law enforcement investigations.     

香港特别行政区的DNA数据库

香港特区的DNA数据库成立于2001年,法例上此数据库直属于香港警务处长,由香港政府化验师代行管理及维护。被定罪人士其罪行最高刑期为监禁7年或以上及任何罪案之疑犯均可被套取其口腔拭子样本,利用美国App lied B iosystem s的试剂进行DNA分型测试。所确立之DNA图谱将上载到由美国FB I(联邦调查局)在2000年免费为香港政府化验所提供的,名为COD IS(Comb ined DNA Index System)的计算机系统;在罪案现场所收集到的不明来历的DNA数据样本亦会被上载到COD IS。被定罪人士及疑犯的DNA图谱会定期与罪案现场的DNA数据在CO-D IS内进行较对以便协助警方调查。本文介绍香港DNA数据库的操作,其中包括统计数据的讨论。     

Comparing Standard and Selective Degradation DNA Extraction Methods: Results from a Field Experiment with Sexual Assault Kits,

A growing number of U.S. cities have large numbers of untested sexual assault kits (SAKs) in police property facilities. Testing older kits and maintaining current case work will be challenging for forensic laboratories, creating a need for more efficient testing methods. Methods: We evaluated selective degradation methods for DNA extraction using actual case work from a sample of previously unsubmitted SAKs in Detroit, Michigan. We randomly assigned 350 kits to either standard or selective degradation testing methods and then compared DNA testing rates and CODIS entry rates between the two groups. Results and conclusions: Continuation‐ratio modeling showed no significant differences, indicating that the selective degradation method had no decrement in performance relative to customary methods. Follow‐up equivalence tests indicated that CODIS entry rates for the two methods could differ by more than ±5%. Selective degradation methods required less personnel time for testing and scientific review than standard testing.     

Developmental Validation of the ANDE 6C System for Rapid DNA Analysis of Forensic Casework and DVI Samples

A developmental validation was performed to demonstrate reliability, reproducibility, and robustness of the ANDE Rapid DNA Identification System for processing of crime scene and disaster victim identification (DVI) samples. A total of 1705 samples were evaluated, including blood, oral epithelial samples from drinking containers, samples on FTA and untreated paper, semen, bone, and soft tissues. This study was conducted to address the FBI’s Quality Assurance Standards on developmental validation and to accumulate data from a sufficient number of unique donors and sample types to meet NDIS submission requirements for acceptance of the ANDE Expert System for casework use. To date, no Expert System has been approved for such samples, but the results of this study demonstrated that the automated Expert System performs similarly to conventional laboratory data analysis. Furthermore, Rapid DNA analysis demonstrated accuracy, precision, resolution, concordance, and reproducibility that were comparable to conventional processing along with appropriate species specificity, limit of detection, performance in the presence of inhibitors. No lane-to-lane or run-to-run contamination was observed, and the system correctly identified the presence of mixtures. Taken together, the ANDE instrument, I-Chip consumable, FlexPlex chemistry (a 27-locus STR assay compatible with all widely used global loci, including the CODIS core 20 loci), and automated Expert System successfully processed and interpreted more than 1200 unique samples with over 99.99% concordant CODIS alleles. This extensive developmental validation data provides support for broad use of the system by agencies and accredited forensic laboratories in single-source suspect-evidence comparisons, local database searches, and DVI.     

A comparative study of two extraction methods routinely used for DNA recovery from simulated post coital samples

In sexual assault cases DNA profiling of spermatozoa can be of critical importance. Most methods use differential extraction of the spermatozoa to separate it from the female component. Here we have compared two commercially available differential extraction methods, the QIAamp^® DNA mini kit (Qiagen) and Differex™ with the DNA IQ^® System (Promega). Simulated postcoital samples were prepared using buccal cells from a female donor and spermatozoa from three male donors. A dilution series ranging from neat semen to a 1:1500 dilution (semen:dH₂O) was prepared and mixed with an equal volume of saliva from a female donor. Extraction efficiency was assessed using DNA concentration measured with NanoDrop 2000 and Quantifiler^® Human DNA Quantification Kit and the profile count of full, partial and mixed DNA profiles generated using SGM Plus and PowerPlex^® ESI 17. Statistical analysis was carried out using Randomisation in R, which is a robust model making no assumption of the distribution of data. Based on the amount of DNA extracted and the types of profiles no significant difference in the performance of the two extraction kits was seen. However, the processing time taken with the Differex™ System was about half than that of the QIAamp^® DNA mini kit and involved fewer liquid transfers.     

Analyzing Approaches to the Backlog of Untested Sexual Assault Kits in the U.S.A.

Motivated by the debate over how to deal with the huge backlog of untested sexual assault kits in the U.S.A., we construct and analyze a mathematical model that predicts the expected number of hits (i.e., a new DNA profile matches a DNA sample in the criminal database) as a function of both the proportion of the backlog that is tested and whether the victim–offender relationship is used to prioritize the kits that are tested. Refining the results in Ref. (Criminol Public Policy, 2016, 15, 555), we use data from Detroit, where government funding was used to process ≈15% of their backlog, to predict that prioritizing stranger kits over nonstranger kits leads to only a small improvement in performance (a 0.034 increase in the normalized area under the curve of the hits vs. proportion of backlog tested curve). Two rough but conservative cost‐benefit analyses—one for testing the entire backlog and a marginal one for testing kits from nonstranger assaults—suggest that testing all sexual assault kits in the backlog is quite cost‐effective: for example, spending ≈$1641 to test a kit averts sexual assaults costing ≈$133,484 on average.     

DNA recovery from different evidences in 300 cases of sexual assault

Almost 60% of the DNA evidences analyzed in our laboratory correspond to sexual assault cases. With the aim to assess the efficiency of the DNA IQ System (Promega) in recovering the perpetrator DNA profile, the statistical analysis of results obtained in 300 casework was performed. In such cases, 850 evidence samples were processed. In 71% of the cases the perpetrator DNA profile was detected in at least one of the submitted casework samples, with a minimum of 13 STRs markers typed using the AmpFlSTR Identifiler PCR amplification kit (Applied Biosystem). When the suspect DNA profile was available, 67% matched with the evidence.With regard to the type of evidence, the best performance corresponded to panties, with more than 70% of success in recovering male profile, whereas the efficiency of vaginal swabs was almost 60%, with a higher incidence of victim/perpetrator mixed profiles.     

Real-time forensic DNA analysis at a crime scene using a portable microchip analyzer

An integrated lab-on-a-chip system has been developed and successfully utilized for real-time forensic short tandem repeat (STR) analysis. The microdevice comprises a 160-nL polymerase chain reaction reactor with an on-chip heater and a temperature sensor for thermal cycling, microvalves for fluidic manipulation, a co-injector for sizing standard injection, and a 7-cm-long separation channel for capillary electrophoretic analysis. A 9-plex autosomal STR typing system consisting of amelogenin and eight combined DNA index system (CODIS) core STR loci has been constructed and optimized for this real-time human identification study. Reproducible STR profiles of control DNA samples are obtained in 2 h and 30 min with ≤0.8 bp allele typing accuracy. The minimal amount of DNA required for a complete DNA profile is 100 copies. To critically evaluate the capabilities of our portable microsystem as well as its compatibility with crime scene investigation processes, real-time STR analyses were carried out at a mock crime scene prepared by the Palm Beach County Sheriff's Office (PBSO). Blood stain sample collection, DNA extraction, and STR analyses on the portable microsystem were conducted in the field, and a successful “mock” CODIS hit was generated on the suspect's sample within 6 h. This demonstration of on-site STR analysis establishes the feasibility of real-time DNA typing to identify the contributor of probative biological evidence at a crime scene and for real-time human identification.     

Internal Validation of the AmpFlSTR Yfiler™ Amplification Kit for Use in Forensic Casework

Abstract:  Y-chromosomal short-tandem repeat (Y-STR) amplification has been used in forensic casework at the Bureau of Criminal Apprehension (BCA) Forensic Science Laboratory since 2003. At that time, two separate amplifications were required to type the SWGDAM recommended loci (DYS19, DYS385a/b, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS438, and DYS439). The Yfiler™ kit coamplifies these loci as well as DYS437, DYS448, DYS456, DYS458, DYS635, and Y GATA H4. The Yfiler™ kit was validated following the internal validations outlined in the SWGDAM revised validation guidelines. Our studies show that 0.125 ng of male DNA will generate a complete 17 locus profile and that as little as 0.06 ng of male DNA yields an average of nine loci. In the male–male mixtures, a complete profile from the minor component was detected up to 1:5 ratio; most of the alleles of the minor component were detected at a 1:10 ratio and more than half the alleles of the minor component were detected at a 1:20 ratio. Complete YSTR profiles were obtained when 500 pg male DNA was mixed with female DNA at ratios up to 1:1000. At ratios of 1:5000 and 1:10,000 (male DNA to female DNA) inhibition of the YSTR amplification was evident. The YSTR results obtained for the adjudicated case samples gave significantly more probative information than the autosomal results. Our studies demonstrate that the Yfiler™ kit is extremely sensitive, does not exhibit cross-reactivity with female DNA, successfully types male DNA in the presence of overwhelming amounts of female DNA and is successful in typing actual forensic samples from adjudicated cases.     

Automation of the Differential Digestion Process of Sexual Assault Evidence,

Sexual assault evidence samples require the use of a specific process known as a differential digestion to separate sperm from nonsperm cells prior to DNA extraction. An automated differential digestion process was developed using a selective degradation technique, which uses DNase I to digest the remaining nonsperm DNA in the sperm fraction. The use of DNase on pristine samples, as well as aged and degraded samples, was assessed to ensure that the quantity and quality of the sperm DNA were not compromised or adversely affected. Samples processed using the selective degradation technique yielded comparable DNA yield and DNA typing data to the conventional differential digestion process. The automated process utilized 96‐well plates for high throughput and incorporated microscope slide preparations for confirmation of sperm. It reduced processing time by about sixfold and was paramount in the elimination of the Oakland Police Department Criminalistics Laboratory's sexual assault kit backlog.     

Why Police “Couldn't or Wouldn't” Submit Sexual Assault Kits for Forensic DNA Testing: A Focal Concerns Theory Analysis of Untested Rape Kits

In jurisdictions throughout United States, thousands of sexual assault kits (SAKs) (also termed “rape kits”) have not been submitted by the police for forensic DNA testing. DNA evidence may be helpful to sexual assault investigations and prosecutions by identifying offenders, revealing serial offenders through DNA matches across cases, and exonerating those who have been wrongly accused, so it is important to understand why police are not utilizing this evidence. In this study, we applied focal concerns theory to understand discretionary practices in rape kit testing. We conducted a three‐year ethnography in one city that had large numbers of untested SAKs—Detroit, Michigan—to understand why thousands of SAKs collected between 1980 and 2009 were never submitted by the police for forensic DNA testing. Drawing upon observational, interview, and archival data, we found that while practical concerns regarding resources available for forensic analysis were clearly a factor, as Detroit did not have the funding or staffing to test all SAKs and investigate all reported rapes, focal concerns regarding victim credibility and victim cooperation were more influential in explaining why rape kits were not tested. Implications for the criminal justice system response to sexual assault and rape kit testing legislation are examined.     

Assessment of the Yfiler® Plus PCR amplification kit for the detection of male DNA in semen-negative sexual assault cases

The ability to detect male epithelial cells deposited during digital penetration or penile penetration without ejaculation is limited by the sensitivity of the Y-STR profiling kit. In this study, the relative profiling success of the Thermofisher Yfiler® Plus kit was compared to its predecessor, AmpFlSTR Yfiler®, for 104 semen-negative sexual assault samples from casework at Forensic Science SA, Adelaide, South Australia. Yfiler Plus generated allele information in 25% more samples than Yfiler and gave a higher recovery of informative alleles in all but two samples where detectable male DNA was present. Where a profile was obtained in both kits, 92% of samples gave a higher percentage of informative loci with Yfiler Plus compared to Yfiler. Yfiler Plus also resolved DNA mixtures in 15 samples as compared to 1 sample with Yfiler. Detection of male DNA with the Quantifiler™ Trio DNA Quantification kit was shown to correlate with a successful profiling outcome with Yfiler Plus. The success of profiling with Yfiler Plus was independent of the time elapsed between the alleged offence and the sample being collected, the type of sexual penetration which occurred, and the anatomical origin of the sample.     

Results of the 2018 Rapid DNA Maturity Assessment

Three commercially available integrated rapid DNA instruments were tested as a part of a rapid DNA maturity assessment in July of 2018. The assessment was conducted with sets of blinded single-source reference samples provided to participants for testing on the individual rapid platforms within their laboratories. The data were returned to the National Institute of Standards and Technology (NIST) for review and analysis. Both FBI-defined automated review (Rapid DNA Analysis) and manual review (Modified Rapid DNA Analysis) of the datasets were conducted to assess the success of genotyping the 20 Combined DNA Index System (CODIS) core STR loci and full profiles generated by the instruments. Genotype results from the multiple platforms, participating laboratories, and STR typing chemistries were combined into a single analysis. The Rapid DNA Analysis resulted in a success rate of 80% for full profiles (85% for the 20 CODIS core loci) with automated analysis. Modified Rapid DNA Analysis resulted in a success rate of 90% for both the CODIS 20 core loci and full profiles (all attempted loci per chemistry). An analysis of the peak height ratios demonstrated that 95% of all heterozygous alleles were above 59% heterozygote balance. For base-pair sizing precision, the precision was below the standard 0.5 bp deviation for both the ANDE 6C System and the RapidHIT 200.