Showing posts with label secondary transfer. Show all posts
Showing posts with label secondary transfer. Show all posts

Monday, December 15, 2014

A reconsideration of the DNA forensics in the Duke lacrosse case


Updated 5/18/2015

Introduction
In the spring of 2006 three players on the Duke lacrosse team were indicted for rape and kidnapping.  The accuser claimed to have been raped by three men at least one of whom ejaculated inside of her body.  Some DNA results emerged in the spring of 2006. Eventually additional evidence was uncovered, and the overwhelming perception that this evidence was exculpatory brought about a major turning point in public opinion prior to the declaration of innocence.  After evidence of his misconduct emerged, the district attorney withdrew from the case, which then fell to the office of the Attorney General of North Carolina to investigate.  After the investigation was complete, the Attorney General declared the three players to be innocent in the spring of 2007, and the criminal matter ended.

The Duke lacrosse case is generally perceived as a highly visible case of prosecutorial misconduct and a rush to judgment by the press and the general public.  However with the 2014 publication of William Cohan’s book The Price of Silence, this view has now been challenged, and Mr. Cohan’s view of the DNA evidence is at odds with many who have commented on the case.  Therefore, a reevaluation of the DNA evidence is warranted on the basis of a general understanding of this case, as well as other cases in which the DNA evidence plays a central role, such as the Knox/Sollecito case, a frequent subject of this blog.

The initial results from the rape kit:  Is the absence of evidence evidence of absence?

The initial results from the NC SBI laboratory did not find semen, blood, or saliva.  Subsequently Y-chromosomal (Y-STR) tests were performed at a private laboratory.  Investigators found no DNA from any Duke lacrosse player in the rape kit items but did find DNA from Mangum’s boyfriend.  DNA from plastic fingernails was also extracted.  Some of the results from the private lab were released in a brief report issued on May 10, 2006, but other results were withheld.  Apart from the plastic fingernails (discussed below), how significant was the lack of DNA?

One blogger posed questions along these lines to several highly qualified experts, including Dan Krane, Elizabeth Johnson, and William Thompson.  Dan Krane responded: “It is worth noting that DNA tests are amazingly sensitive (DNA profiles can be generated from as little material as that left behind in a fingerprint) and Y-STR tests have the potential of determining a male’s DNA profile even when a female’s DNA is present in hundreds or thousands of times greater quantities. Scientists are always wary of asserting that the absence of evidence is not proof of absence but it certainly is reasonable to expect to find a rapist’s DNA associated with a victim when the victim presents herself to investigators within hours of an attack and when she has not bathed, the rapist did not use a condom and ejaculation occurred.”
  These experts gave thoughtful, nuanced replies. The consensus view seemed to be that the degree to which the absence of evidence was evidence of absence lies in the particulars of each case.

What then are the particulars of the Duke lacrosse case?  A site at Reed College lists the following guidelines for preserving evidence of a possible sexual assault:

 “Do not shower or douche
Try not to urinate. Urinating may reduce the ability to detect “date rape” drugs
 If there was oral contact, do not smoke, eat, or brush teeth
 Do not change clothes. If you have already changed your clothes, place them in a paper bag (plastic may destroy evidence) If you haven’t changed, keep the original clothes on and bring an extra set to wear home from the hospital
Go to a hospital with the capability of providing a SAFE exam and request the exam. The cost of a SAFE examination is paid for from a state fund.”

The alleged victim in the Duke lacrosse case was taken into custody shortly after the incident and did not have the opportunity to shower or to change clothes.  Therefore, the chances of preserving evidence would have been high, if an actual assault had occurred.  William Anderson wrote, “Furthermore, no one — no one — who inspected the room (and police gave that bathroom a thorough inspection, taking swabs everywhere and testing for DNA) found any evidence of bleach products having been recently used. They found no DNA of Mangum, and none of [Collin] Finnerty and [Reade] Seligmann, and nothing else that demonstrated that either of those two young men had ever been in that bathroom.”  Parenthetically, Taylor and Johnson reported that no fingerprints belonging to Mr. Finnerty were found in the bathroom (Until Proven Innocent, p. 183).

Unfortunately, the question of whether or not the accuser said that the victims used condoms has been not entirely without controversy.  Stuart Taylor and KC Johnson wrote (Until Proven Innocent, p. 327), “In her report from the night of the alleged attack, [SANE nurse Tara] Levicy had written without qualification that Mangum had said repeatedly…the rapists had used no condoms and that she was sure that at least one had ejaculated—in her mouth.  But on January 10 [2007, nine months later], according to [Linwood] Wilson, Levicy stated that Mangum “said ‘no’ but wasn’t really sure” whether her attackers had used condoms.”  Even if one finds Ms. Levicy to be credible about Ms. Mangum’s lack of certainty (which I do not), DNA may be transferred by simple physical contact, as is borne out in studies of simulated strangulation.  In addition, perspiration contains DNA, and sebaceous fluid is a probable source of touch DNA, according to a study by Vecchiotti, Filippini and their coworkers.  Thus even when a condom is used during a sexual assault, there may be transfer of DNA.

Asked whether or not the use of a condom could have been detected, Elizabeth Johnson responded, “Testing for these substances is not typically done, despite what you see on CSI. There has been some research done re spermicides on condoms, but none of this is done as part of a typical test and validated methods for casework aren’t there yet.”  Of course, even if condom use were detected, it would not differentiate between consensual and nonconsensual sexual activity.

The lack of body fluids is absence of evidence in its own right.  It also weakens the DNA evidence, meaning that a given DNA profile found in the absence of a particular fluid might have arisen from any biological tissue or fluid.  Peter Gill describes framework or hierarchy of levels (Misleading DNA Evidence, p. 19) at which DNA evidence can be evaluated:
1.     The sub-source level refers to the strength of evidence of the DNA profile itself.
2.     The source level is an evaluation of the strength of the DNA profile if it can be associated with a particular body fluid, such as semen or blood
3.     The activity level associates the DNA with the crime itself, e.g. sexual assault.
4.     The highest level deals with the ultimate issue of guilt/innocence.

Professor Gill’s book gives examples of cases in which a forensic scientist attempted to claim a higher level for the evidence than was warranted, based upon principles of forensics and probability.  The DNA of the rape kit is sub-source DNA, in this categorization.

The lack of DNA from the accused coupled with the lack of body fluids and the lack of other evidence in the bathroom is strongly exculpatory.  Whether or not it constitutes “proof of innocence” cannot be determined without defining the term.  However, I would weight this lack of evidence more heavily that eyewitness testimony, even if the latter came from a credible witness, owing to the well-known problems with eyewitness testimony.  Ms. Mangum’s credibility is questionable on a number of grounds.

The plastic fingernails in the trashcan
Ms. Mangum’s painted fingernails were placed in the trashcan of the bathroom and the players pointed them out to the police investigators days later.  David Evans could not be excluded as a donor.  The summary of conclusions from the office of the Attorney General of North Carolina stated on p. 12, "First, statistically, the chance of randomly selecting an individual from the population that could be included in this sample would be approximately 1 in 1000." With respect to the autosomal (standard DNA) profile, David Evans (along with about 2% of the population) cannot be excluded as a donor.  Likewise in the Y-chromosomal (YSTR) testing, David Evans cannot be excluded as a donor.  Although the summary report did not explain the calculations, the value of 1 in 1000 probably derives from combining the data from the two types of DNA testing.

The author of a 2014 book on the case (The Price of Silence), William Cohan clearly thought that the DNA was highly incriminating, discussing it on pp. 277-278 and pp. 325-326, among other places.   In the final chapter, Denouement, he returned to the subject on p. 602:  “What remains unresolved is, if in fact it was David Evans’s DNA on Mangum’s red plastic fingernails, how did it get there?”

DNA evidence beneath real fingernails typically is probative, not necessarily conclusive, evidence against a defendant.  Studies have shown that mixtures of DNA under fingernails are seen about 5% of the time (Gill, pp. 43-46).  However, there is less to the DNA from the plastic fingernails than meets the eye.  Taylor and Johnson wrote (Until Proven Innocent, p. 221), “And it would later become clear…that the DNA that might (or might not) have come from Evans had not necessarily been lodged “under” a plastic fingernail.  Rather, the SBI had extracted and mixed together all of the DNA taken from all surfaces of the three used plastic fingernails found in the wastebasket in Evans’ bathroom.”  Moreover, a case in Britain involving a taxi driver, David Butler suggests some additional caveats.  His DNA was apparently found on the fingernails of a murdered woman Anne Marie Foy.  He was convicted partially on the basis of the DNA evidence but later released.  A plausible theory is that he handled money which later carried his DNA to the victim.  There are not many forensic details that have been made public, but a few things can be gleaned from the information that is available.  One is that Mr. Butler's skin was unusually flaky.  Two is Hannah Barnes' report that "The victim was also wearing a glitter nail polish, which proved particularly attractive to dirt - and DNA." 

The defense correctly noted that David Evans was not uniquely identified in either of the tests.  The defense’s position was that if David Evans were a donor of the DNA, then there existed a plausible route for innocent DNA deposition, namely secondary transfer from materials in trashcan, such as dental floss and tissue paper.  Defense attorney Joseph Cheshire, "Any expert and any person in the world will tell you that your [own] DNA is in your bathroom." (Cohan, p. 242).  Saliva is a good source of DNA, for example.  The office of the attorney general of North Carolina accepted the possibility of secondary transfer as a reasonable explanation.  In the summary document pertaining to this case, the authors wrote: “Third, to the extent that Evans’s DNA could not be excluded, the SBI experts confirmed that the DNA could easily have been transferred to the fingernails from other materials in the trash can.”

It is uncontroversial that the presence of DNA does not indicate the time or manner of its deposition.  This cuts both ways: one cannot be certain that the DNA on the plastic fingernails arrived by secondary transfer, but neither can one rule it out.  Secondary transfer in DNA forensics is well established, and tertiary transfer has been observed under some circumstances.  Neither the amount of DNA nor the completeness of the DNA profile is a reliable guide as to whether the DNA arrived by primary transfer or secondary transfer.  Joe Minor and Suzanna Ryan have written several well-researched and readable articles that cover the subjects of DNA found on touched objects and secondary transfer.  Review articles by Van Oorshot and colleagues and by Meakin and Jamieson in academic forensic journals may be consulted for more detailed information.

Shedding and transferring DNA is perhaps more common than the general public appreciates.  Leslie Prey wrote, "We all shed DNA, leaving traces of our identity practically everywhere we go. Forensic scientists use DNA left behind on cigarette butts, phones, handles, keyboards, cups, and numerous other objects, not to mention the genetic content found in drops of bodily fluid, like blood and semen (Van Oorschot & Jones, 1997)."  The issues of shedding and transferring DNA were perhaps not very important when DNA profiling was in its infancy.  However, the introduction of techniques based on the polymerase chain reaction (PCR) meant that much smaller amounts of DNA are needed now than in the first generation of DNA profiling.  This is because the DNA is increased in amount (amplified) many times over prior to the analysis.  The more recent development of low copy number DNA profiling means that still smaller amounts of DNA can be typed, less than a dozen cells.

Although secondary transfer from the waste in the trashcan is highly plausible, primary or secondary transfer via the players themselves is also possible.
  Joe Minor wrote, “On one occasion, I swabbed my own hand after handshakes at a social function to determine the presence of other epithelial cells.  The laboratory was able to obtain a mixture of my DNA as well as two other individuals.”  Suzanna Ryan wrote, “One study performed by Lowe, et al., was designed to highlight a ‘worst case’ scenario and involved two individuals. The first was determined to be a poor shedder and the second a good shedder. These two shook hands for one minute. The poor shedder had washed their hands immediately prior to the experiment whereas the good shedder had not. After shaking hands the poor shedder held a sterile plastic tube for 10 seconds. The tube was then swabbed and tested for the presence of DNA. This experiment was performed on two sets of good shedder/poor shedder pairs. Surprisingly, in one of the pairs, only the good shedder's DNA was obtained from the plastic tube, with no evidence of a mixture including the poor shedder!”

  Results from Vecchiotti’s laboratory published in 2014 are consistent with this result.  Suppose that David Evans shook hands with Ms. Mangum when she arrived. That or a number of other events, such as handling money, might also transfer DNA.

  As a team captain and resident of the house, Mr. Evans might have shaken hands with Ms. Mangum when she entered or touched money that she later handled.

As persuasive as these arguments are however, one even more powerful argument may also be offered in Mr. Evans’ defense using the tables that Professor Giannelli presented in the chapter “DNA Profiling” within the book Race to Injustice.  At least two other males contributed DNA to the sample (not lacrosse players nor other men who attended the party).  In the YSTR profile ten of sixteen loci (locations within the Y chromosome) show the presence of three contributors and two more loci show the presence of at least two contributors.  In the autosomal profile some loci (places within other chromosomes) clearly show that at least three individuals contributed DNA.  This is very problematic for the prosecution.  If one acknowledges that this DNA arrived in a way unrelated to a sexual assault, then how is it possible to exclude the possibility that Mr. Evans’s DNA also arrived innocently?

By itself, the presence of what may be Mr. Evans’ DNA on the fingernails seems inculpatory; in the context of the other profiles and given the strong possibility of secondary transfer by multiple mechanisms, it is close to meaningless as evidence.  Paul Giannelli summed up the DNA on the plastic fingernails: “There was some probative value--but not much.”  Followers of the Knox-Sollecito case will recognize that the presence of additional male DNA donors is basically the same situation that exists with Mr. Sollecito’s Y-chromosomal profile that is associated with the bra clasp.  The Gary Leiterman case is perhaps the starkest example of this conundrum for the prosecution; the additional profile belongs to someone who could not possibly have been present at the crime scene.

Two more issues deserve attention, one of which is the difference in how the SBI versus DNA security interpreted the mixture in the autosomal DNA profile.  Taylor and Johnson wrote (Until Proven Innocent, p, 221), “The SBI had reported to Nifong that there were ‘multiple contributors’ to the DNA mixture but that ‘no conclusion [could] be rendered’ as to whether any of this fingernail DNA was even a possible partial match with any of the players…Based on the same testing process that had led the state lab to find no basis for any conclusion at all, DNA Security offered a singularly weak conclusion: that 98% of the male population could be ‘excluded as a contributor to the mixture’ and that David Evans was in the other 2 percent.”  Section 21C of the motion to compel discovery also quotes the same portion of the SBI report, and footnote 70 emphasized that the SBI had said “that no conclusion ‘could be rendered as to the contribution of DNA profiles from the buccal swabs of the suspects’ in the minor profile BS 4522.” 

The defense was rightly concerned about this apparent discrepancy.  It is possible that DNA Security used a suspect-centered analysis, meaning that they analyzed the mixture with Evans’s reference profile in hand.  However in “Painting the target around the matching profile: the Texas sharpshooter fallacy in forensic DNA interpretation,” William Thompson observed, “In the absence of clear standards for ‘inclusion’ and ‘exclusion’, different experts evaluating the same evidence may reach different conclusions: one may conclude that a particular suspect is ‘included’, while another concludes that the same suspect is ‘excluded’.”  Indeed, Dror and Hampikian have shown that the same mixed DNA profile may be interpreted differently by different groups of analysts.  Unfortunately the interpretation of DNA mixtures is presently still a somewhat subjective process; therefore, the difference between the two labs may have an innocent explanation.  However, another take-home message from this case is that the defense should re-analyze DNA mixtures from items of evidence.

Finally, Dr. Meehan’s own DNA may have contaminated an item of evidence.  Such contamination events are not uncommon.  The ABA’s standard 4.1a states in part:

“(ix) reports of laboratory contamination and other laboratory problems affecting testing procedures or results relevant to the evaluation of the procedures and test results obtained in the case and corrective actions taken in response…”

If there were a contamination event, Dr. Meehan’s laboratory should have reported it.  However, laboratories do not always do so.  In the Adam Scott and Gary Leiterman cases, the laboratories simply ignored evidence of DNA contamination in the negative controls (negative controls are experiments which should have no DNA in them).  There are also instances in which negative controls were faked in some way, as noted by William Thompson in “Tarnish on the gold standard.”  The reasons to fake having performed the negative controls include covering up a contamination event and increasing laboratory throughput.  Given the existence of DNA contamination and of instances of samples being mixed up or mislabeled in some way, it is surprising and disappointing that anyone opposes complete disclosure of the raw data, contamination logs, standard operating procedures, and any other pertinent records.

The DNA evidence released in the fall
On 27 October 2006 Nifong released some 1800 pages of material to the defense, but still did not include a complete report.  Although he lacked training in this area, defense attorney Brad Bannon did an extensive study of the material and was able to glean the essential information that the Y- chromosomal DNA of four unidentified men (men who were not at the party or Mangum’s boyfriend) had been found in the items of evidence from the rape kit.  When this information become public knowledge, it was a major turning point in the case, one that helped to shift public opinion in favor of the three accused students.

These profiles are important for at least three reasons.  One is that the finding of DNA serves as evidence that the tests were working, in other words the results functioned as a positive control.  The technique of amplifying DNA using the polymerase chain reaction (which is central to modern profiling) is susceptible to inhibitors of DNA polymerase, the enzyme that copies the DNA.  The lack of a DNA profile might in principle, arise from the presence of inhibitors, as opposed to the absence of DNA.  Of course, it is likely that the lab ran positive controls, but this sort of information might not come to light unless a DNA expert had reviewed the case for the defense.  Two is that Ms. Mangum’s sexual activity provides an explanation for the edema noted in her examination.  Three is that Ms. Mangum had claimed no sexual activity for a week, and these results raised doubts about her credibility, which would have been a central issue if the case had come to trial.

Using the information that they had obtained a month and a half earlier, the defense teams jointly crafted a motion to compel discovery and filed it on 13 December 2006.  Besides the information gleaned above, the defense teams learned that DNA Security had tested more items than it had disclosed in its report:

There are significant gaps in the item sequence numbers…The worksheet shows that the DNA extraction was performed n July 12, 2006, on that item, as well as items labeled “16074C” and “16075C,” which are nowhere otherwise identified in the DNA Security materials.  But on this worksheet itself, under the “name” column, someone typed “Bobby” and “Owen.”  “Quantification Worksheet Q-181”75 and “PCR Worksheet Number A-464”76 reflect quantification & amplification work on those three extractions the following day, July 14, 2006.  “Analysis Worksheet Number G-592” reflects analysis of those extractions the following day, July 14, 2006.  But nowhere in the materials do the DNA profiles generated from those analyses appear.  The phenomenon repeats itself throughout the underlying materials from DNA security.77

Students of the Knox/Sollecito case are familiar with this kind of withholding of evidence, except on a larger scale; there are many samples for which DNA quantification was performed but for which no electropherogram (loosely speaking, an electropherogram is a picture of the sizes of the DNA fragments) was produced.

DNA Security’s standard operating procedures with respect to its reports were shown to be in marked contrast with its conduct in this case.
52.  Included in the materials provided to the Defendants from DNA Security are the laboratory’s standard operating procedures, which include guidelines for drafting “Reports and Documentation” of analyses performed by the lab in each case:

Reports shall include:
·      Case identifier.
·      Description of evidence examined.
·      Description of methodology.
·      Name of each DNA locus analyzed.
·      Results for each DNA test.
·      Interpretative statement of conclusions…

That the results for each test should be part of the report is very clear.  Yet Dr. Meehan chose to disregard this standard for reasons that are open to speculation.  Perhaps he acted as he did because he wished to establish a business relationship with the office of the district attorney or because of the publicity that working on a high-profile case would bring his firm.  The defense motion also noted, “the troubling effect of its limited scope of reporting is that it allowed DNA security to produce a report in this case that avoided disclosure of exculpatory evidence, either in the form of potential contamination noted in its testing, or as it relates to the discovery of DNA from multiple male sources…”  It is difficult to overstate how different the actual results are from the results as portrayed in the incomplete report from DNA Security produced on 12 May 2006.

DNA laboratory reports and the discovery of DNA evidence
The American Bar Association’s model rules make several important points regarding DNA evidence, one of which also insists that the reports be complete.  Standard 3.3a states, “(a) A summary of all DNA testing and data interpretation should be recorded promptly in a report.”  Standard 4.1a states in part, “(xi) material or information within the prosecutor’s possession or control, including laboratory information or material, that would tend to negate the guilt of the defendant or reduce the punishment of the defendant.”

Robert Mosteller discussed Nifong’s transgressions: “Nifong had an ethical duty under Rule 3.8(d) to provide the exculpatory information in a timely fashion, which is not explained further by the rule, but lacks any suggestion that prolonged unjustified delay is authorized.”  Kenneth Williams concurred, writing, “By withholding exculpatory evidence from the defense, Nifong violated the U.S. Constitution, the laws of North Carolina, and Rule 3.8d of the North Carolina Rules of Professional Conduct.  Specifically, the State Bar found that ‘Nifong did not make timely disclosure to the defense of all evidence or information known to him that tended to negate the guilt of the accused’ and that he ‘failed to make a reasonably diligent effort to comply with a legally proper discovery request.’”

Professor Paul Giannelli commented on the incomplete laboratory reports, “In any event, no attorney should have to search through the haystack for the exculpatory needle.  A laboratory report should be comprehensive and include a section specifying the limitations of the technique used in the analysis.  The report should also be comprehensible to laypersons.” (emphasis mine)  Professor Giannelli noted that Dr. Brian Meehan also failed to meet the standards for reports laid out by the American Society of Crime Directors/Laboratory Accreditation, even though his laboratory relied upon this organization for accreditation.

Conclusions
The circumstances surrounding the absence of evidence coupled with the presence of DNA from other men is persuasive evidence of absence; no one who attended the party sexually assaulted Ms. Mangum.  The DNA that may be from David Evans on the plastic fingernailsis very weak evidence against him; in isolation it would raise the possibility of his participation in a sexual assault.  However, there are plausible alternative explanations for how his DNA arrived; therefore, if this were the only piece of evidence in the case, it would not come anywhere close to the threshold of reasonable doubt.  In the context of the absence of Mr. Evans DNA in the rape kit and the presence of other men’s DNA on the plastic fingernails, it loses almost all significance.

Regrettably, the withholding of evidence is a common theme in miscarriages of justice; however, two factors were not in Mr. Nifong’s favor.  In addition to North Carolina’s open discovery law, the defense would also have been helped by the norms of discovery of DNA evidence as set forth in the ABA standards, which would have allowed expert review of the data in its raw form.  Without any discovery of the exculpatory DNA evidence, the case might have played out much differently, especially if Mr. Nifong had been able to escape censure.  If Mr. Nifong had brought the case to trial, it is difficult to predict how a jury would weigh the fingernail DNA evidence against potential alibi witnesses for Mr. Evans (Mr. Finnerty and Mr. Seligman had electronic alibis).  Juries have occasionally ignored strong alibi evidence in favor of weak or questionable evidence, as in the cases of Jonathan Fleming and Russ Faria.  Fortunately, the state's investigators came to the conclusion that Mr. Nifong should have, and Attorney general Roy Cooper ended the matter.

Update 5/18/2015
When Brad Bannon was preparing to defend David Evans, he read John Butler's textbook on DNA profiling.  His cross-examination of Dr. Brian Meehan was one of the pivotal moments of the case.  For the newest edition of his three-volume textbook Dr. Butler asked Mr. Bannon to comment on aspects of how a defense attorney would challenge DNA evidence.  Mr. Bannon replied in part, "Did the lab follow acceptable standards of DNA analysis?  Did it follow its own protocols?  Is the lab applying those standards and protocols consistently or selectively?  For example, why do you call a peak below 150 RFU as a true allele for one purpose, or in one case, but not for another?  If there are such internal inconsistencies, do they usually inure to the benefit of one side's theory of the case?  If so, is that evidence of bias?"

Mr. Bannon's point about consistency of threshold values for peaks is similar to one found in the textbook An Introduction to Forensic DNA analysis, 2nd ed. (Rudin, N. and Inman, K., CRC Press 2002, p. 121) states, “It is important to have some predetermined limit to distinguish what is signal and what is noise.”  Without a clear guideline, a scientist may make choices that benefit his or her preferred hypothesis, even subconsciously.

A partial list of authors and references
 William Anderson “Duke:  Why the DNA mattered.”  William Anderson teaches economics at Frostburg State University in Maryland.  He was one of the first bloggers to call attention to the many problems in the Duke lacrosse case.

William Cohan, The Price of Silence, Scribner, 2014.  Mr. Cohan is a writer on business affairs.

Paul Giannelli, “DNA Profiling” in Race to Injustice (2009), Michael Siegel, ed.
Paul Giannelli is Albert J. Weatherhead III and Richard W. Weatherhead Professor of Law at Case Western University in Ohio.  He was the Reporter for the American Bar Association Criminal Justice Standards on DNA evidence, and he specializes in the area of scientific evidence.  William Thompson explained, "The Reporter for an ABA Standards Group or Study Committee is the person whose job it is to take down and 'report' the findings, conclusions and determinations of the group or committee.  It is the reporter who prepares the first draft of the report and of the annotations and notes that accompany a report.  The Reporter usually works under the direction of a committee chair, but has independent responsibility for getting details right."


Peter Gill, Misleading DNA Evidence, Academic Press, 2014.  Peter Gill is Professor of Forensic Genetics at The University of Oslo, Norway and is a professor at the Norwegian Institute of Public Health.  He is chair of the DNA commission of the International Society of Forensic Genetics, and he has published more than 180 peer-reviewed papers.

Georgina Meakin and Alan Jamieson, “DNA transfer: Review and implications for casework,” Forensic Science International: Genetics 7 (2013) 434–443.  Professor Jamieson is the head of the Forensic Institute in Glasgow, Scotland.

Robert P. Mosteller, Exculpatory Evidence, Ethics, and the Road to the Disbarment of Mike Nifong: The Critical Importance of Full Open-File Discovery, 15 George Mason Law Review 257-318 (2008).  Robert Mosteller is J. Dickson Phillips Distinguished Professor of Law at The University of North Carolina, Chapel Hill.  He teaches Evidence, Criminal Procedure Investigation, Constitutional Criminal Procedure and co-directs the Trial Advocacy Program.

Joe Minor, “Touch DNA: From the Crime Scene to the Crime Laboratory,” Forensic Magazine 4/12/13.  Joe Minor teaches forensic science and is a forensic DNA consultant.

Suzanna Ryan “Touch DNA Analysis: Using The Literature To Help Answer Some Common Questions.”  Forensic Magazine.  Suzanna Ryan is a contributor to law enforcement and forensic magazines, and she is a consultant and expert witness.

Stuart Taylor and KC Johnson, Until Proven Innocent, St. Martin’s Press, 2007.
KC Johnson hosted Durham in Wonderland, a blog that was a major source of information related to the case.  Stuart Taylor is a freelance journalist and a contributing editor for the National Journal, specializing in law.  He is a nonresident fellow of the Brookings Institution.

William Thompson, Painting the target around the matching profile: the Texas sharpshooter fallacy in forensic DNA interpretation.” Law, Probability and Risk (2009) 8, 257-276.

William Thompson, “Tarnish on the Gold Standard.”  William Thompson is a professor in the Department of Criminology, Law and Society at the University of California, Irvine.  His areas of expertise include forensics and human judgment and decision-making.

Van Oorshot R.A., Ballantyne, K.N., and Mitchell, R.J.  “Forensic trace DNA:  a review.  Investigative Genetics 1 (2010) 1:14. doi: 10.1186/2041-2223-1-14.  Roland Van Oorshot is a member of the Forensic Services Department of the Victoria Police, Victoria, Australia.

Kenneth Williams “An Examination of the District Attorney’s Alleged Unethical Conduct,” in Race to Injustice (2009), Michael Siegel, ed.  Kenneth Williams is a member of the faculty of the South Texas College of Law in Houston, TX.

Monday, August 19, 2013

DNA does fly, and it also transfers quite easily


Part 37 in the Knox/Sollecito case

Update (11/7/2013)
Poy and Van Oorshot wrote, "To further help evaluate the above finding swabs were taken from gloves worn whilst examining a heavily soiled dress during routine casework examination. A significant amount of DNA was retrieved which exhibited a genetic profile that matched that of samples taken from the exhibit."  This is direct evidence that gloves can transfer DNA.

Introduction
The issues of possible DNA contamination and proper versus improper handing of evidence are at the center of the forensic evidence in the trials of Amanda Knox and Raffaele Sollecito.  This entry will examine two aspects of DNA forensics, dealing with airborne DNA and the need to change gloves when handling a fresh item of evidence.  Both kinds of potential problems need to be recognized and addressed in order to minimize the chances of contamination.

The problem of airborne DNA
Barbie Nadeau wrote, “…the defense claims that the crime scene was badly compromised during the collection of evidence.  Alberto Intini, head of Italy’s national forensic team, disagrees. On the stand, he defended the forensics work and stressed that the crime scene had not been contaminated, especially under cross examination when the defense lawyers tried and failed to prove otherwise. ‘DNA does not fly around like pollen,’ he said…” Ms. Nadeau left her readers with a misimpression.  Later in the trial the defense returned to this issue.  “’DNA does not have wings, but it flies,’ [defense expert witness Sarah] Gino cautioned. ‘In a laboratory where hundreds of samples are examined, the risk of contamination exists and should be taken into consideration.’”

Dr. Intini’s view is seriously in error, as can be ascertained by a number of lines of evidence.  A paper on DNA in fossils (BioTechniques 38:569-575, April 2005) notes, “These molecules are easily spread via aerosol transport. One aerosol droplet can contain many more DNA molecules than one gram of fossil material.” In a discussion of how to collect DNA evidence Dick Warrington advised, “Next, you can prevent contamination by wearing a mask, since you want to avoid coughing and sneezing around the evidence you are processing.”

pipettors and pipet tips
Pipettes are devices used to deliver small volumes of liquids.  They are used frequently when the polymerase chain reaction (PCR) technique is used to amplify tiny amounts of DNA.  For that reason the use of aerosol barrier pipette tips is routine in labs engaging in PCR.  An application note on pipetting explained, “Cross-contamination occurs if improper pipetting causes splashes or drips. Even if pipettors are handled properly, aerosols can be generated that contain DNA molecules and can contaminate the pipettor and subsequent pipet products. This demands increasing efforts to reduce the penetration of contaminants by means of filter tips.”  Gilson, a manufacturer of pipettes, supplies a technical bulletin that reads in part: “For example, if a technician in a crime lab performs PCR on a blood sample, cross-contamination between samples could result in an erroneous incrimination, even if the technician changes pipette tips between samples. A few blood cells could volatilize in the pipette shaft, stick to the plastic of the pipette, and then get ejected into the next test sample. Modern laboratories have taken account of this fact and are devoting tremendous efforts to avoid this problem through the use of filter tips."

laboratory traffic
A number of forensic guidelines are in place to protect against aerosol DNA from one source or another.  For example,  “A ‘one-way traffic’ rule is also observed in the laboratory, once the technician has entered the PCR or the post-PCR rooms, they are not allowed to return to the extraction or pre-PCR rooms until the next day or a complete cloth changing in order to prevent contamination by aerosol particles.”  The problems only get more serious in the low template region of analysis.  Keith Bedford said, “The way I am speaking at the moment, we could probably detect DNA on this pad in front of me.”

laboratory design
Laboratories are constructed in a way to minimize the possibility of contamination due to air flow.  In the article Setting Up a PCR Laboratory.” Theodore E. Mifflin discussed how the design of the laboratory can minimize the chances of contamination: “Air handling. For extremely high-performance PCR laboratories that will be involved with detecting very-low-prevalence DNA or RNA molecules (e.g., infectious disease agents in clinical samples), additional measures may be necessary to prevent contamination from the air being recirculated between the pre- and post-PCR laboratories. In this case, the air handlers need to be separate and the air pressure individually adjusted in each laboratory. In the pre-PCR laboratory, there should be a slight positive pressure compared to the air in the connecting hallway. The post-PCR laboratory, in contrast, should be at slightly reduced pressure to pull air in from the outside and thereby prevent escape of amplicons from the completed PCR samples being analyzed inside the lab (Fig. 2). Finally, the air handlers for the pre- and post-PCR laboratories need to be connected to separate air ducts, and each must lead to a separate location for exhaust.”  Dr. Mifflin’s main focus is pathology, but his points about low levels of DNA are germane to low template DNA forensics in that both situations use PCR to amplify very small quantities of DNA. 

What the police did and did not do with respect to changing gloves
When the forensic police collected evidence at Ms. Kercher’s flat, they did not change their gloves frequently.  One can observe in a series of photographs a failure to change gloves over several minutes and several evidence samples.  Barbie Nadeau reported, “Sollecito’s attorney, Giulia Bongiorno, stopped the crime scene video several times to point out errors. For example, Stefanoni testified that she had changed gloves according to official investigation procedures, but Bongiorno stopped the crime scene video twice to show that Stefanoni’s bracelet and the fold of her glove were exactly the same before and after the time she claimed to have changed gloves.”  Ms. Nadeau recounts the same incident on p. 133 of Angel Face, her book on the murder of Meredith Kercher.

Ms. Stefanoni’s views on when gloves should be changed are found in the English translation of the Massei report.  On p. 203 Massei wrote that Stefanoni "specified" that gloves were changed "every time an object was touched that was particularly soaked with blood, and when it was obvious that the gloves would be soiled;"  Based on pages 204-205, she appears to believe  that the presence of a liquid is necessary to bring about contamination by touch.

What forensic experts say about changing gloves
On page 38 of John Butler's 2005 textbook Forensic DNA Typing, he wrote, "Use clean latex gloves for collecting each item of evidence. Gloves should be changed between handling of different items of evidence."  Dick Warrington is the author of some articles in Forensic Magazine and is employed by a company which makes equipment for crime scene investigations.  He wrote, “If you pick up one piece of evidence and then pick up another piece of evidence you can transfer evidence from the first item to the second item. You can avoid this kind of cross-contamination if you remember to change your gloves before handling each piece of evidence.”  He also advised, “Put on gloves, use gloves, change gloves. Do that every time you touch a piece of evidence. Likewise, use disposable tweezers, scalpels, etc. Change these each time they are used, as well.”  Orchid Cellmark’s guidelines for collecting DNA evidence read in part, “Use clean latex gloves for collecting each item of evidence. It is recommended the gloves be changed between the collection of each item of evidence.”

Conclusions
The need to use fresh gloves when handling a new piece of evidence is utterly noncontroversial.  Yet it is obvious from video of the collection, that the forensic police did not change gloves frequently.  Their failure to follow the consensus view of what is good practice might have contaminated the bra clasp and might have led to mixed DNA samples elsewhere in the flat.  The fact that airborne DNA can compromise an experiment is likewise the consensus view of practitioners of PCR.  Dr. Intini’s ignorance of the dangers of aerosol DNA shake one’s confidence in the ability of a forensic lab under his supervision to combat the dangers of airborne DNA contamination even in routine DNA profiling, let alone low template profiling.

Tuesday, July 13, 2010

DNA transfer in Strangulation

Part XIX in the Knox/Sollecito case

Update 1, 22 August 2010

I communicated with an anonymous forensic nurse about their domestic violence program. If an alleged victim complained of an attempted strangulation, complained of a partner’s grabbing their arm, or showed bruising or redness, they have been swabbing that area for DNA, for two years. Positive results in at least one case helped to convict someone. This information confirms the reasonableness of swabbing Meredith’s body in appropriate places, although the bruise on the nape of Meredith’s neck may have been the result of her being thrown against the wall (as some have theorized). We do not know whether swabbing the bruised areas was done or not.
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PM Guiliano Mignini’s reconstruction of the murder of Meredith Kercher was shown to the jury in the form of an animated video. In his speculation Amanda Knox grabbed Meredith Kercher by the throat and slammed her against a wall. The video superimposed actual shots of Meredith’s bruises with Amanda’s animated hand to imply that Amanda’s action produced the bruises. Later, Rudy Guede and Raffaele Sollecito held Meredith’s arms back, and Amanda stabs her (Barbie Nadeau, Angel Face, p. 160). In forensics professor Carlo Torre’s reconstruction of the crime, the single assailant grabbed Meredith by the throat and stabbed her. The area under Meredith’s chin and her nape were bruised; the former bruising was the result of the knife and the latter when the assailant put Meredith down.

Although Mignini’s theory of the crime suggests a much longer amount of contact between the assailants and victim than does Torre’s, both imply contact. None of Amanda Knox’s DNA was found in Meredith’s bedroom or on her body. Raffaele Sollecito’s DNA was found on a bra clasp (although the defense contests this piece of evidence) but not the bra itself. Rudy Guede’s DNA was found in several places, including on the sleeve of Meredith’s sweatshirt. Let us examine some forensic DNA studies to see if they shed any light on this tragedy.

The study by Wiegand and Kleiber gives a case study in which DNA evidence was collected 48 hours after a strangulation. They wrote, “Strangulation marks were clearly visible on the neck of the victim. Epithelial cells could be removed from the neck of the victim using separate cotton swabs for the left and the right side of the neck. Only the swab from the right side could be typed and included the pattern of the suspect (Fig. 3), a result which corresponded to the autopsy findings (the right neck side showed a higher intensity of bleeding in the muscles than the left side indicating a more intensive pressure against the right side). Altogether clear results could be obtained using four STRs (TH01, VWA, FGA, CD4) demonstrating the high utility and sensitivity of the method described.” These authors also conducted simulated strangulations, and they reported a success rate of better than 70%.

The simulated strangulation study in 2002 by Rutty used both SGMplus and LCN amplification. The simulated strangulation experiment was done with periods between the force and the sampling were 1, 5, 10, 15, 30, and 60 min, 2, 3, 4, 5, 6, 7, 8, 24, and 48 h and 3, 4, 5 and 10 days. When the author used SGMplus, he observed a full profile of the offender 7 out of 29 times, and always in the presence of the victim’s profile. When the author used LCN all 17 experiments yielded offender profiles, with the majority being partial profiles. Dr. Rutty wrote, “Of the test neck swabs, 19 yielded positive amplification results using SGMplus, 12 showed a victim-only profile and 7 a victim and offender profile with a full offender profile detectable up to 6 h after contact. When LCN was used (17 tests) all showed the offender to be present for all time periods i.e. up to 10 days. In the majority of cases it was a partial offender profile with the majority of the amplification result being a full victim profile.”

Dr. Rutty stated that “When considering the apparent time periods of DNA survival, passive transfer of the offender’s DNA onto the victim’s neck could also explain the presence of offender DNA several days after contact.” This study did not break down the results by time periods between simulated strangulation and DNA collection; therefore, it is difficult to draw firm conclusions based solely on the data presented.

A 2008 study by Graham and Rutty reexamined the question of DNA transfer to and from strangulation victims with an emphasis on innocent DNA transfers that might deposit DNA on the neck of a victim. In 24% of samples collected showed nonself DNA on the simulated victim from third party sources. The authors believe that such DNA transfers might confuse an investigation.

Finally, it might be helpful to return to the subject of primary and secondary DNA deposition. The study by Lowe and coworkers in 2002 looked at DNA transfer from a good DNA shedder to a poor DNA shedder to an object. By definition transfer was secondary from the good shedder, and transfer was primary from the poor shedder. When mixtures were observed, secondary transfer from the good shedder provided the major component, not primary transfer from the poor shedder. This study illustrates the dictum that one generally cannot infer the mechanism of how the DNA was deposited from the DNA itself. Along with the study by Graham and Rutty, this work suggests caution in the interpretation of nonself DNA on the body of the victim of strangulation. A similar caution should also be applied in the interpretation of DNA on the bra clasp.

In addition to the case study discussed by Wiegand and Kleiber, there are a small number of news reports of strangulations that mention DNA. One is from greater Detroit, and another is from Chicago. However, these articles do not specify the place on the body that was tested for DNA. In addition to the possibility that DNA was collected from the neck, it is possible that the victim’s fingernails contained the perpetrator’s skin cells or vice versa.

There are no reports that ILE found anyone’s DNA on Meredith’s bare wrists or her neck, and it is unclear whether or not the forensic police swabbed for DNA in these areas. It is also unclear whether some areas on the neck would have been free enough of Meredith’s blood to allow swabbing for the assailant’s DNA. Nevertheless it is difficult to see why the forensic police should not have swabbed the nape of Meredith’s neck or her wrists. If they did and found nothing, it would be strongly exculpatory, although it might fall short of proof of innocence due to uncertainties over collecting enough DNA or collecting it quickly enough after the murder. If the forensic police failed to swab these areas, it would suggest that they did not do as thorough a job as one would wish.

Bibliography
P. Wiegand and M. Kleiber, “DNA typing of epithelial cells after strangulation,” International Journal of Legal Medicine (1997) 110 :181–183. abstract

G. N. Rutty, “An investigation into the transference and survivability
of human DNA following simulated manual strangulation
with consideration of the problem of third party contamination,” International Journal of Legal Medicine (2002) 116 :170–173.
abstract

A. Lowe, C. Murray, J. Whitaker, G. Tully, P. Gill, “The propensity of individuals to deposit DNA and secondary transfer of low level DNA from individuals to inert surfaces.” Forensic Science International (2002) 129(1):25-34. abstract

E. A. M. Graham and G.N. Rutty, “Investigation into ‘normal’ background DNA on adult necks: implications for DNA profiling of manual strangulation victims.” Journal of Forensic Science, (2008) 53(5):1074-82. abstract