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ESTABLISHING PENETRATION IN SEXUAL ASSAULT CASES

Jennifer Gentile Long, JD, Viktoria Kristiansson, JD, and Charlene Whitman-Barr, JD1 INTRODUCTION

Criminal acts of sexual violence generally fall into three categories: exposure, contact, and penetration. While prosecu¬tors introduce evidence to establish the statutory elements at trial, defense strategies focus on targeting any vulnera¬bility in that evidence. Challenges to identification, consent, and attacks on victim credibility remain the most common defense tactics in any sexual violence crime. Where the charged offense includes an element of penetration, defenses may also include specific challenges to the prosecution's ability to prove that penetration occurred. If the prosecution is unable to prove the element of penetration beyond a reasonable doubt, the accused will be acquitted or convicted of a less serious offense.2

This STRATEGIES in Brief explains the legal requirements for establishing penetration in sexual assault prosecutions and offers strategies for effectively identifying, evaluating, and presenting evidence of penetration. First, it summarizes the categories of criminal sex offense statutes and outlines the legal requirements to establish penetration. Second, it pro¬vides strategies to prepare for and try sexual assault cases involving penetration. Third, it identifies and offers guidance for responding to common defense challenges to establishing penetration in sexual assault cases.

KNOW YOUR LAW: THE LEGAL DEFINITION OF PENETRATION

The difference between the common understanding of the term “penetration” and the legal definition of the term “pen¬etration” can create confusion for victims, witnesses, and even criminal justice professionals. The consequences of this confusion can result in inaccurate investigative reports, failures to record or preserve critical evidence or statements, or the mischarging of a case. This section, therefore, discusses the legal elements of penetration among the 58 jurisdictions of the United States.3

Sexual penetration crimes include the penetration of the vagina,4 anus, or mouth5 by the penis or other body part, and also include the penetration of the vagina or anus, or, in rare cases, the mouth, by an object. Penetration with an object is included in all jurisdictions’ sexual assault statutes except Louisiana's and American Samoa's.6 Penetration of a victim's mouth with an inanimate object is criminalized as a sexual offense only in Alabama, New Hampshire, and under the Uniform Code of Military Justice.7 Since penetration of the mouth by an object is not specifically covered by the sexual assault statutes in most jurisdictions, charges for such conduct can be filed under other assault-related crimes or under a provision for a sexually-motivated felony (e.g., certain categories of assault, strangulation, or kidnapping).8

Vaginal penetration occurs, under the law, when the penis, other body part, or object enters the vulva or between the labia majora, which is the outermost part of the female genital organ.9 Anal penetration occurs, under the law, when the penis, other body part, or object enters the anal opening;10 at least one court has determined that penetration of the buttocks is insufficient to establish anal penetration under its jurisdiction's definition of sexual intercourse.11 Oral penetration occurs when the penis, other body part, or object enters the lips of a victim's mouth,12 and has also been found to occur by the act of licking a penis.13 In some jurisdictions, the courts have held that penetration can be proven if it occurs through clothing.14

 

 

 

No jurisdictions require more than “slight penetration,” but not all statutes use the term “slight” in their statutes.15 In states that have statutes that do not specifically enumerate the requirement that penetration need only be “slight,” one must consult the relevant case law for this element; treatises also provide examples and further guidance.16 No jurisdic­tions’ penetration statutes require ejaculation.17

Evaluating Evidence and Strengthening the Case with Corroboration

A sexual assault charge can be proven beyond a reasonable doubt solely with credible victim testimony; no corroboration is required in order to establish the elements. Some jurisdictions, however, have statutes or case law that, under limited circumstances, such as when a victim is mentally incapacitated during the assault, require corroborative evidence.18 Al­though corroboration is not required, where available, it strengthens the victim’s testimony given the persistence of rape myths, which cause juries to search for reasons to doubt victim testimony in sexual assault cases.19 Research concerning sexual violence supports some common explanations for the routine lack of “traditional” forms of corroborating evidence in these cases. For example, since these crimes are typically committed in solitude, there are rarely direct eyewitnesses.20 In addition, because the trauma resulting from these crimes may impact the willingness or ability of victims to report,21 physical evidence may no longer be available or observable. Further, many sexual assaults do not result in physical injury to a victim’s genitalia. Where injury does exist, medical examinations and records may not observe or document them, due to factors including the examiner’s level of training and experience, whether enhanced observation equipment22 was used, or if there was a gap in time between the incident and the examination.23 Even in cases where traditional corrobo­ration is present, defense attorneys will still challenge evidence of penetration.24 These prosecutorial issues demonstrate the need for the thorough and accurate identification, review, and documentation of any and all corroborating evidence in sexual assault cases.

Victim Testimony

When evaluating a case, it is important to remember that victims may be impacted by the effects of trauma when they report sexual assault crimes, which in turn may affect their ability to communicate with authorities, family, and friends.25 Their communications may be nonlinear, piecemeal, disjointed, or inconsistent, and, when combined with an inaccurate or incomplete understanding of the legal definition of penetration, may result in various professionals’ documentation or categorization of events that fail to capture all of the facts or criminal elements. A trauma-informed approach to screen­ing and victim interviews can improve accuracy while providing important support to victims, a critical component of maintaining victim engagement with the system.26 Prosecutors using a trauma-informed approach can have the thor­ough discussions with victims necessary to ensure the elements can be proven, while reducing any collateral negative impact that participation in the prosecutorial process can have on the victim.

In many cases, a victim’s testimony is the best way to establish the element of penetration. Evidence of sensory details, such as what a victim heard, saw, felt, tasted, and even smelled, is highly relevant evidence. The specificity that this cat­egory of evidence provides strengthens the credibility of any witness’s testimony, but can prove particularly powerful when the ability to prove a charge rests with the victim’s testimony and her/his credibility. Regardless of whether an event occurred before, during, or after penetration, the event and its associated sensory details may be relevant to estab­lishing penetration.

 

 

 

 

PRACTICE TIPS

The below list is not exhaustive but provides examples of topics that can be addressed in the interview and direct examination of the victim:

•A perpetrator’s threats, commands, or other words can indicate an intent to penetrate the victim.

•The exposure of the perpetrator’s penis, finger, or penetrating object can corroborate the intent and ability to perform penetration.

•The physical feelings that the victim experienced can also demonstrate penetration, including, if applicable, the perpetrator’s failed attempts to penetrate the victim prior to achieving penetration.

•If the offender penetrated the victim’s mouth, testimony regarding a gagging sensation, taste, or the feeling caused by the penis or object against the victim’s mouth or clenched teeth can be sufficient to establish penetration achieved between the victim’s lips.

•The perpetrator’s removal of clothing (perpetrator’s and/or victim’s) can demonstrate the intent and ability to penetrate the victim.

•The use of a condom or questions about the victim’s use of prophylactic can indicate an intent to penetrate.

•The victim’s state of mind (e.g., fear of sexually transmitted infection (STI) or pregnancy, or fear of the perpetrator) can corroborate the perpetrator’s act of penetration.

•The victim’s contraction of an STI or post-assault pregnancy.

•Leakage of semen, blood, or other fluid.

•Post-assault injury or discomfort.

•Post-assault pain during urination or bowel movement.

Additional sources of evidence can support the victim’s testimony of penetration and related facts. In cases where the victim is unable to testify to the assault, e.g., if she was unconscious or incapacitated during the assault, or where she is unable to communicate at the time of trial, those additional sources of evidence may be necessary to establish penetra­tion. A discussion of different types of evidence follows below.

Witness Testimony

The act of penetration may have occurred in a public place or a place easily accessible by other persons. Where a rape or sexual assault occurs in the presence of one or more witnesses, there may be eyewitness testimony that is relevant to the act of penetration.27 Witnesses may have directly observed the penetration of the anus, vagina, or mouth. Even where witnesses do not actually observe penetration by a body part or object, they may have witnessed other events that are relevant, such as the perpetrator’s use of a condom, the perpetrator’s or the victim’s own statements, or statements by bystanders or co-conspirators, which can directly or circumstantially prove penetration occurred.28 Witnesses may have observed all or part of the assault, and perhaps for just a short period of time. The act of penetration can take just one second, and the opportunity for the perpetrator to commit the act may require little time and limited privacy.

 

 

 

 

Defendant Statements

Where defendants choose to make statements to investigators, even their self-serving statements may help the prosecu­tion establish penetration. Examples include defendants who allege the victim consented to the penetration, allege the defendant acted “by mistake,” or offer other noncriminal explanations for their actions. In their statements, perpetrators may lock themselves into a specific defense (e.g., consent), in which they admit penetration, and therefore establish or corroborate a required element. Even where such statements do not discuss penetration specifically, they may still corroborate penetration by implication. A defendant’s statement can bolster a victim’s description of penetration by corroborating words spoken by the victim during the incident that are relevant to establishing penetration, such as excla­mations of pain, while providing an alternative explanation for them. Defendant statements may also include discussions about the victim’s stated concern about pregnancy or sexually transmitted infections (STIs) or the perpetrator’s lack of condom use. Finally, defendants alleging consent may describe their own concerns about pregnancy or STIs. When prosecutors focus on the more obvious defense strategies, they may overlook or minimize corroborating evidence in defendants’ own statements.

Prosecutors should carefully weigh the question of whether to introduce a defendant’s self-serving statement. Although the statement may help prove the element of penetration, its introduction may provide critical support for the defense theory without necessitating the defendant’s testimony. The last thing that the prosecution wants to do is establish the element of penetration but lose the case because the prosecution handed the entire defense strategy to defense counsel by introducing an out-of-court statement that the defense would not otherwise have been able to introduce.

Where the defendant takes the stand, prosecutors can, through cross-examination, corroborate the act of penetration, thereby foreclosing any defense attempt to argue that the element of penetration was not proved beyond a reasonable doubt. In cases where the defendant does not testify, a prosecutor may choose, under some circumstances, to seek to in­troduce defendant’s statements, where permissible under the rules of hearsay, through the victim, or potentially through other witnesses to support other evidence of penetration.29

Digital Evidence

The availability of video and photographic evidence of sexual assaults has increased along with the explosion of smart­phones, apps, surveillance video, and social media. If such evidence is present in a case, it may provide important corrob­oration of penetration. However, even where a video clearly appears to show penetration, a defense attorney may still argue that the fact finder cannot be certain that the tape depicts penetration due to the camera angle, distance from the subject, or other factors. Prosecutors should carefully review such video evidence to determine its potential for inves­tigative leads and weight at trial.

For more information go to EVAWINTL.org

College freshman charged with rape says he was just re-enacting ‘Fifty Shades of Grey*

By Kirsten Anderson

CHICAGO, February 24, 2015 (LifeSiteNews.com) -- A University of Illinois-Chicago freshman is being held on $500,000 bail for a violent sexual assault he claims was a consensual re-enactment of the bestselling novel and hit film “Fifty Shades of Grey.”

Mohammad Hossain, 19, is accused of binding, gagging, beating, and raping a fellow UIC student in his dorm room last Saturday evening during what may have begun as a consensual physical encounter.

According to Assistant State's Attorney Sarah Karr, the alleged victim had been intimate with Hossain before, and willingly went to his dorm room. After she undressed, Hossain then bound her arms and legs with belts, gagged her with a necktie, covered her eyes with a knit cap, and began whipping her with a belt.

At that point, Karr said, the alleged victim “began shaking her head and crying” and begged Hossain to stop. Instead, Hossain continued to beat her with the belt and his fists.

Eventually, the alleged victim managed to free herself from Hossain’s restraints, but Hossain pinned her down with his arms and sexually assaulted her. Soon after the assault, Hossain’s roommate returned, but Hossain held the door closed to block him from entering the room. When the woman finally escaped, she called police, who arrested Hossain. Hossain reportedly told them he and the alleged victim were re-enacting scenes from "Fifty Shades of Grey."

As described by prosecutors, the alleged assault does bear similarities to “Fifty Shades of Grey.” Hossain allegedly gagged his victim with a necktie - an item that has become an iconic symbol of the popular series about bondage and sexual domination, gracing the cover of the first novel. The alleged whipping of his victim with a belt mirrors the climactic scene in the first novel and currently playing film, in which sadistic male lead Christian Grey finally alienates his prey-turned-paramour by beating her so violently that she leaves him.

The Chicago Tribune reported that after Hossain’s bail was set at $500,000, the student leader and athlete - he serves as an ambassador to the alumni association and participates on the triathlon team - appeared stunned, his mouth wide open as he was escorted back to his holding cell. Asked by the judge what would possess an otherwise respected young man to “let a movie persuade him to do something like this,” Hossain’s defense attorney said her client believed the encounter was consensual.

Sign a petition to boycott Fifty Shades of Grey here.

Detection of Male DNA in the Vaginal Cavity After Digital Penetration Using Y-Chromosome Short Tandem Repeats

Kayla R. Sween, MS, Lawrence A. Quarino, PhD, and Janine M. Kishbaugh, MS ABSTRACT

In this study, useful genetic information from male donors was obtained on vaginal swabs taken from female volunteers after male digital vaginal penetration in a time frame relevant to a sexual assault investigation. Vag­inal swabs were collected from eight volunteers at intervals of 1, 6, 12, 24, and 72 hours after digital vaginal penetration. DNA was extracted from collected swabs and subsequently genotyped using a commercially available Y-chromosome short tandem repeats (Y-STR) multiplex kit. Fifty-eight vaginal swabs were collected and analyzed in the study. Composite Y-STR profiles from all combined volunteers showed that 85% of all pos­sible alleles were detected at the 1 -hour interval, 77% of all possible alleles were detected at the 6-hour inter­val, 73% of all possible alleles were detected at the 12-hour interval, 66% of all possible alleles were detected at the 24-hour time interval, and 71% of all possible alleles were detected at 72 hours after digital vaginal pen­etration. Results indicate that a viable possibility exists that probative Y-STR profiles, useful for investigative purposes, can be obtained from vaginal swabs taken from subjects exposed to digital penetration at time in­tervals up to 72 hours postpenetration.

KEYWORDS:

digital penetration; DNA profiling; sexual assault evidence collection; Y-STRs

 

 

 

 

T

 

he purpose of this study is to determine the viability of obtaining Y-chromosomal DNA profiles consis­tent with those of a male donor on vaginal swabs taken from female volunteers at various time intervals after male digital vaginal penetration. Successfully obtaining male DNA profiles may encourage jurisdictions to develop the policy of collecting vaginal swabs from victims when this type of assault is alleged.

In 2011, the Federal Bureau of Investigation (FBI) Criminal Justice Advisory Policy Board voted to change the definition

Author Affiliations: Forensic Science Program, Cedar Crest College. All financial support and instrumentation were provided by Cedar Crest College except for reagents and instrumentation for DNA quantitation of swab extracts, which were provided by Thermo Fisher Scientific in Frederick, MD.

The authors declare no conflict of interest.

Correspondence: Janine M. Kishbaugh, MS, Forensic Science Program, Cedar Crest College, 100 College Drive, Allentown, PA 18104. E-mail: jmkish@cedarcrest.edu.

Received September 08, 2014; accepted for publication December 03, 2014.

Copyright © 2015 International Association of Forensic Nurses DOI: 10.1097/JFN.0000000000000056

Journal of Forensic Nursing of rape in the Uniform Crime Reporting Summary Reporting System because of objections from many women’s rights activist groups. The new FBI definition of rape expanded the 82-year-old Uniform Crime Reporting definition of rape from forcible penile-vaginal penetration to “penetration, no matter how slight, of the vagina or anus with any body part or object, or oral penetration by a sex organ of another per­son without the consent of the victim” (Women’s Law Project, 2011). The FBI collects statistics regarding rape annually; in 2011, there were approximately 83,500 rapes of adult women reported to law enforcement agencies. In addition, over 80,000 female children are sexually assaulted in the United States annually (Bonnar-Kidd, 2010). Since the change in the definition of rape, it is important to consider the collection of additional evidence when sexual assaults are reported or when any sexual-assault-related incident occurs. According to the new FBI definition of rape, digi­tal penetration is also considered a form of rape necessitat­ing the need to collect physical evidence from the victim if digital penetration is alleged.

Digital penetration commonly results in genital trauma (Grossin et al., 2003). Although genital trauma is identified through visual inspection, histological techniques, and

www.journalforensicnursing.com 33

colposcopy (Sommers, 2007), the presence of evidence from the act of digital penetration is not investigated. Digital pen­etration is most frequendy reported in young children who are sexually abused, although the report of these assaults is less frequent than assaults on adult women (Finkel, 1998; Grossin et al., 2003).

It is currently unknown how the vaginal environment affects the detection and presence of male epithelial cells as a function of time after male epithelial cell deposition. No studies have been conducted on the detection of male DNA in the vaginal cavity after digital penetration. A pre­vious study pertaining to digital penetration only investi­gated the persistence of female epithelial cells under male fingernails up to 72 hours after digital penetration (Flanagan &C McAlister, 2011). This indicates that there is transfer and persistence of epithelial cells after digital penetration and sug­gests that male DNA could also persist and be detected from the vaginal cavity.

Sexual assaults involving penile-vaginal penetration nor­mally result in a DNA mixture of male and female DNA when using autosomal short tandem repeats (STRs; Butler, 2005). However, in some cases of mixed DNA profiles, the quantity of male DNA is too low to obtain a meaningful male profile using autosomal DNA analysis. A previous study successfully utilized Y-chromosome STRs (Y-STR) profiling to analyze vaginal swabs collected from sexually assaulted female victims where spermatozoa were not detected on the swabs and autosomal male DNA typing results were not obtained. The maximum time frame for successful Y-STR DNA typing was found to be 48 hours after the assault and was likely because of low quantities of undetected sperm cell or male epithelial cells (Sibille et al., 2002).

Y-STR profiles were also obtained in female subjects after extended postcoital intervals (Mayntz-Press & Ballantyne, 2008; Quarino & Kishbaugh, 2012). Usable Y-STR pro­files were obtained in vaginal swabs at postcoital intervals as long as 8 days (Quarino & Kishbaugh, 2012).

In this study, vaginal swabs were taken from volunteers at various intervals after digital penetration and subsequently examined for Y-chromosomal DNA using a commercially available Y-STR multiplex to ascertain whether a male- specific DNA profile from the digital penetration can be de­tected in the vaginal cavity.

Methods

Approval to Work With Human Subjects The study was reviewed by the institutional review board of Cedar Crest College who approved this project for use with human subjects. Approval for use of human subjects was granted only after information regarding volunteer re­cruitment procedures, methods of sample collection, and protections of confidentiality were adequately described to the institutional review board of Cedar Crest College.

34 www.journalforensicnursing.com

Informed consent procedures were carried out. In addition, the institutional review board ensured that any possible risks associated with the collection of samples in this study were explained to the volunteers and that they had a right to withdraw from the study at any time. The names of human subjects used in this study are known only to the authors.

Sample Collection

Eight couples were asked to participate in this study without regard to age of the subjects. No specific criteria for volun­teer recruitment were used. The participants in the study did not have any known chronic medical conditions or acute illnesses. Couples who did not use condoms during sexual activity were asked to abstain from sexual activity for at least 14 days before digital penetration. Those couples who typically used condoms during sexual activity began the study after abstaining from sexual activity for 7 days. Af­ter digital penetration was performed for 5 minutes, the vaginal cavity was swabbed at one of the following time intervals: 1, 6,12,24, or 72 hours after penetration. Cou­ples also abstained from sexual activity during each of the requested time intervals. Instructions on sample collection were provided by the researchers. Immediately before dig­ital penetration, four control vaginal swabs were collected by the woman. Two swabs were simultaneously inserted into the vaginal cavity and then removed. Two additional swabs were then inserted, creating two sets of two control swabs. After digital penetration, the vaginal cavity was swabbed at one of the time intervals mentioned above by the woman in the same manner as the control swabs. Upon collection of the samples, the two sets of two swabs were randomly designated either A or B. There was no correlation between the letter and the order of collection.

After collection, swabs were reinserted into the original swab packaging, cotton tip facing outward protruding from the open area of the packaging, and allowed to air dry. Once dried, the swabs were placed into manila envelopes to keep each collection separate from the others. Collection of bio­logical samples into a paper medium is recommended to pre­vent microbial growth from occurring (Butler, 2005). Swab sets A and B for both controls and samples were packaged separately. Four more trials were performed to collect samples at the remaining intervals in the same fashion.

During the tested time intervals, female volunteers en­gaged in normal activity, but none performed feminine wash. Specific information about bathing and showering during the tested time intervals was not collected.

Buccal swabs were collected from the male and female participants to serve as typing controls for the study. Only five couples finished all of the collection time intervals. Fifty- eight samples were collected and analyzed in the study (two samples each from 29 intervals).

Volume 11 • Number 1 • January-March 2015

Contamination Prevention Procedures in the Laboratory

Gloves were worn at all times when working with biological samples or handling materials for DNA-related procedures and were changed when handling different samples. All work surfaces and materials (e.g., tubes, tube racks) were cleaned with 10% domestic bleach followed by 70% ethanol before all analytical procedures. Before DNA extraction, quanti­tative polymerase chain reaction (PCR), conventional PCR, and capillary electrophoresis, all supplies (e.g., mo­lecular grade water, pipettes, microcentrifuge tubes) were irradiated in an Ultraviolet Stratalinker 2400 Crosslinker (Stratagene, La Jolla, CA) for 75 minutes to minimize con­tamination of samples.

Analytical Methods

All analytical methods to be described are typically used in forensic science laboratories and are consistent with accepted analytical procedures as described by the FBI (2011a). Swab sets A and B were analyzed separately.

DNA Extraction

DNA from each swab set was extracted in an AirCIean600 PCR Workstation (AirCleanR Systems, Raleigh, NC) using the DNA IQ System (Promega, Madison, WI). The proce­dure for DNA IQ is described by the manufacturer (Promega Corporation, 2013). The final sample was eluted in 100 mL of DNA IQ Elution Buffer.

DNA Quantitation

DNA quantitation of exemplar male buccal swabs was per­formed by real-time PCR with a Rotor-Gene 6000 (Qiagen, Germantown, MD) using the Alu-based method employing SYBR Green for detection as described (Nicklas & Buel, 2003). Vaginal DNA samples were quantified for male- specific DNA using the Quantifiler Trio DNA Quantification Kit (Life Technologies, Carlsbad, CA) and a 7500 Real-Time PCR System (Life Technologies, Carlsbad, CA) according to the manufacturer's guidelines (Life Technologies, 2014).

DNA Amplification and Genotyping

DNA samples were amplified and subsequently genotyped using the PowerPlex Y23 kit (Promega, Madison, WI). The protocol was followed according to the manufacturer’s guidelines (Promega Corporation, 2014), and the specific parameters for genotyping were obtained from those re­ported (Pasino, Caratti, & Robino, 2012). Total reaction volume of 25 ml was used for all amplification reactions. The samples were amplified with a Life Technologies Veriti 96 Well Thermal Cycler (Life Technologies, Carlsbad, CA) and analyzed by capillary electrophoresis using an Applied Biosystems Prism 310 Genetic Analyzer (Life Technologies, Carlsbad, CA) with an injection time of 5 seconds. DNA amplicons were sized using CC5ILS 500 (Promega, Madison,

Journal of Forensic Nursing

WI; standard size) and genotyped using Genemapper ID-X software version 1.0 (Life Technologies, Carlsbad, CA). An analytical threshold of 50 relative fluorescence units (RFU) was set for allele identification. An analytical threshold of 50 RFU is typically used in forensic biology laboratories. Be­low the 50 RFU threshold, users run the risk that artifactual peaks may be misidentified as true DNA alleles and allelic drop-in and dropout because of stochastic effects (only one allele detected in a heterozygote genotype because of pref­erential amplification of the detected allele) at low DNA concentrations (typically below the 50 RFU threshold) may become problematic (Bregu et al., 2012). The analysis method was created according to the manufacturer’s guidelines (Promega Corporation, 2014).

DNA Concentration and Purification

Swabs from each couple yielding incomplete profiles were subsequendy concentrated using Amicon Ultra 0.5-mL 100K filters (Millipore, Billerica, MA). Concentration of DNA samples is routine in forensic science laboratories when the amount of human DNA is very low (below the recommended amount of DNA needed for amplification in commercial forensic DNA genotyping kits) or when initial testing re­sults in incomplete profiles. The remaining extract (approx­imately 80 pL) was applied to the filter, and manufacturer’s specifications were followed (Millipore Corporation, 2009). Concentrated samples were reamplified and genotyped using the described procedures. The remaining amplified product was then purified using a MinElute PCR Purification Kit (Qiagen, Germantown, MD) according to manufacturer’s guidelines (Qiagen Sample Assays and Technologies, 2008) and reinjected on the genetic analyzer with no change in electrophoretic parameters. Extracts that yielded full pro­files did not undergo concentration and purification.

Results Data Interpretation

Interpretation of data in this study is consistent with typical guidelines employed in forensic science laboratories and does not deviate from accepted practice as described by the FBI (2011b).

Male-specific DNA Quantitation

With the exception of two test swab extracts, all tested ex­tracts produced DNA quantities below 1 ng of male DNA at the amplification input volume of 17.5 mL. Two 1-hour post-digital-penetration swabs yielded male-specific DNA quantities of 0.0608 and 0.0528 ng/mL, respectively. Be­cause all samples had input volumes of 17.5 mL regardless of quantitation value, 1.18 ng of male-specific DNA was added from the former sample, and 1.02 ng of male- specific DNA was added from the latter sample to their re­spective amplification reactions. This only slightly exceeds the manufacturer recommendation (Promega Corporation,

www.journalforensicnursing.com 35

2014) of adding up to 1 ng of male DNA. These 2 samples, however, produced full Y-STR profiles and thus were unaf­fected by the slight increase in input male DNA.

Individual Swab Sets

Five of the 8 couples in the study produced at least 1 full profile in an individual swab set. Not surprisingly, the trend in the amount of alleles obtained in individual swab sets generally decreased with increasing time interval although there is a slight increase from 24 to 72 hours (see Table 1). Of the 58 swabs collected in the study, 55 underwent con­centration and purification. The 3 swabs that did not yielded complete profiles with the initial extracted sample.

At 1 hour after digital penetration, 2 swab sets were collected from seven couples. Before concentration and pu­rification (listed as “Original” in Table 1), at least 1 set of swabs from 3 of the couples produced full Y-STR profiles (23 alleles). Test swabs from 2 other couples yielded profiles containing 19 alleles, one couple yielded 11 alleles, and one couple yielded eight alleles. With concentration, 2 additional couples (the 2 couples who produced 19 alleles with the orig­inal extract) produced full Y-STR profiles. Results from swabs from 2 of the couples who underwent both concentration and purification were relatively unchanged. Five of the 7 couples (71 %) were therefore able to produce full profiles or nearly full profiles (defined as profiles with 20-22 alleles) 1 hour after digital penetration.

At 6 hours after digital penetration, two swab sets were collected from 6 couples. Before concentration and purifi­cation, swab sets from 2 couples yielded full profiles. No more than 6 alleles were detected in any of the tested swabs from the other 4 couples. With concentration and purification, the 4 couples did see an increase in the number of alleles with one couple producing a maximum of 20 alleles. Three of the 6 couples (50%) were therefore able to produce full profiles or nearly full profiles 6 hours after digital penetration.

At 12 hours after digital penetration, two swab sets were collected from 6 couples for 12 swab sets. Before concentration and purification, no swab set yielded a full or nearly full profile with the maximal number of alleles for all couples combined ranging from 4 to 18. Five of the 6 couples did increase the number of alleles detected after concentration and purification ranging from 10 to 20 (two couples were able to produce 20 alleles). Two of the 6 couples (33 %) were therefore able to produce full profiles or nearly full profiles 12 hours after digital penetration.

At 24 hours after digital penetration, two swab sets were collected from five couples for 10 swab sets. Before con­centration and purification, 1 couple was able to produce a nearly full profile (22 alleles), whereas the maximum num­ber of alleles from the other couples ranged from 0 to 10. The couple who yielded 22 alleles was able to produce a full 23-allele profile after concentration. Concentration and purification did increase the number of alleles in three

36 www.journalforensicnursing.com of the 4 other couples resulting in profiles ranging from 8 to 12 alleles. One of the five couples (20%) was therefore able to produce full profiles or nearly full profiles 24 hours after digital penetration.

At 72 hours after digital penetration, 2 swab sets were collected from 5 couples for 10 swab sets. Before concentra­tion and purification, no swab set yielded a full or nearly full profile with the maximal number of alleles for all cou­ples combined ranging from 0 to 19. Concentration and purification did increase the number of alleles in 4 of the

5  couples, whereas 1 couple showed a significant decrease in the number of alleles (19 alleles before concentration,

5  alleles after concentration, and 8 alleles after purification). Two of the 4 couples who showed an allele increase pro­duced profiles of 20 and 21 alleles, respectively. The elec- tropherogram of the Y-STR DNA profile from the couple who yielded 21 alleles after concentration and purification is found in Figure 1. The other 2 couples yielded only a maximum of 7 alleles after concentration and purification. Two of the 5 couples (40%) were therefore able to pro­duce full profiles or nearly full profiles 72 hours after digital penetration.

None of the control vaginal swabs in the study yielded male DNA.

Composite DNA Profiles

Composite profiles for each couple at each time interval were generated based on the presence of an allele obtained with the original extract, concentrated extract, or concen­trated and purified amplification product in either set of swabs (see Table 1).

At 1 hour after digital penetration, five of the 7 cou­ples had full composite profiles (23 alleles), whereas 2 couples had respective partial composite profiles of 11 and 13 alleles. Of all possible alleles, 85% were detected for all combined couples at the 1-hour interval.

At 6 hours after digital penetration, 1 couple had a full composite profile, 2 couples had nearly full composite pro­files (20-22 alleles), and 3 couples had respective partial composite profiles of 11, 12, and 16 alleles. Of all possible alleles, 77% were detected for all combined couples at the 6-hour interval.

At 12 hours after digital penetration, 3 of the 6 cou­ples had nearly full composite profiles, and 3 couples had re­spective partial composite profiles of 12,12, and 16 alleles. Of all possible alleles, 73% were detected for all combined couples at the 12-hour interval.

At 24 hours after digital penetration, one couple yielded a full profile, whereas 4 couples produced respective partial composite profiles of 9,14,16, and 16 alleles. Of all possible alleles, 66% were detected for all combined couples at the 24-hour interval.

Finally, at the 72-hour postpenetration interval, 1 couple had a full composite profile, 2 couples produced a nearly

Volume 11 • Number 1 • January-March 2015

 

TABLE 1. Number of Alleles Obtained From the Original, Concentrated, and Purified Samples for Swab Sets A and B From Each Couple at All Time Intervals

 

1 hour

6 hours

12 hours

24 hours

72 hours

A

B

A

B

A

B

A

B

A

B

Couple 1

Original

19

19

6

3

0

4

0

10

19

5

 

Concentrated

23

22

4

2

6

0

9

0

5

1

 

Purified

23

23

9

7

6

10

11

5

4

8

 

Composite

23

11

12

14

21

Couple 2

Original

19

10

3

23

8

6

0

5

16

12

 

Concentrated

23

13

12

23

8

12

5

4

18

10

 

Purified

23

17

14

0

13

20

0

12

21

20

 

Composite

23

23

20

16

23

Couple 3

Original

8

0

0

1

6

0

0

0

0

7

 

Concentrated

9

0

11

2

1

0

8

0

4

5

 

Purified

9

0

14

11

11

0

8

0

7

7

 

Composite

11

16

12

9

10

Couple 4

Original

11

2

6

0

15

6

10

3

3

1

 

Concentrated

11

0

12

1

0

2

7

2

12

2

 

Purified

6

2

12

6

0

5

9

9

20

9

 

Composite

13

12

16

16

21

Couple 5

Original

23

23

3

5

NC

NC

NC

NC

NC

NC

 

Concentrated

NT

22

4

11

NC

NC

NC

NC

NC

NC

 

Purified

NT

22

6

20

NC

NC

NC

NC

NC

NC

 

Composite

23

21

NA

NA

NA

Couple 6

Original

NC

NC

NC

NC

7

2

NC

NC

NC

NC

 

Concentrated

NC

NC

NC

NC

19

0

NC

NC

NC

NC

 

Purified

NC

NC

NC

NC

20

1

NC

NC

NC

NC

 

Composite

NA

NA

20

NA

NA

Couple 7

Original

23

17

10

23

18

0

22

19

0

0

 

Concentrated

NT

21

0

23

7

0

23

16

2

0

 

Purified

NT

22

0

23

12

0

23

20

7

0

 

Composite

23

23

21

21

 

Couple 8

Original

23

23

NC

NC

NC

NC

NC

NC

NC

NC

 

Concentrated

NT

23

NC

NC

NC

NC

NC

NC

NC

NC

 

Purified

NT

23

NC

NC

NC

NC

NC

NC

NC

NC