30 results on '"Castella V"'
Search Results
2. Shedding light on the relative DNA contribution of two persons handling the same object
- Author
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Oldoni, F., Castella, V., and Hall, D.
- Published
- 2016
- Full Text
- View/download PDF
3. A new approach for the separation of spermatozoa from other cell types in forensically relevant samples
- Author
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Grosjean, F. and Castella, V.
- Published
- 2015
- Full Text
- View/download PDF
4. Waddlia chondrophila, a Chlamydia-related bacterium, has a negative impact on human spermatozoa
- Author
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Baud, D, Vulliemoz, N, Ammerdorffer, A, Gyger, J, Greub, G, Castella, V, and Stojanov, M
- Published
- 2018
- Full Text
- View/download PDF
5. A Logical Framework for Forensic DNA Interpretation
- Author
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Hicks, T., Buckleton, J., Castella, V., Evett, I., and Jackson, G.
- Subjects
Genetics (clinical) ,Genetics ,DNA ,LR ,activity issues ,evaluative ,forensic ,investigative ,principles of interpretation ,propositions ,reporting ,transfer - Abstract
The forensic community has devoted much effort over the last decades to the development of a logical framework for forensic interpretation, which is essential for the safe administration of justice. We review the research and guidelines that have been published and provide examples of how to implement them in casework. After a discussion on uncertainty in the criminal trial and the roles that the DNA scientist may take, we present the principles of interpretation for evaluative reporting. We show how their application helps to avoid a common fallacy and present strategies that DNA scientists can apply so that they do not transpose the conditional. We then discuss the hierarchy of propositions and explain why it is considered a fundamental concept for the evaluation of biological results and the differences between assessing results given propositions that are at the source level or the activity level. We show the importance of pre-assessment, especially when the questions relate to the alleged activities, and when transfer and persistence need to be considered by the scientists to guide the court. We conclude with a discussion on statement writing and testimony. This provides guidance on how DNA scientists can report in a balanced, transparent, and logical way.
- Published
- 2022
6. Exploring the relative DNA contribution of first and second object’s users on mock touch DNA mixtures
- Author
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Oldoni, F., Castella, V., and Hall, D.
- Published
- 2015
- Full Text
- View/download PDF
7. Waddlia chondrophila, a Chlamydia-related bacterium, has a negative impact on human spermatozoa
- Author
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Baud, D., Vulliemoz, N., Ammerdorffer, A., Gyger, J., Greub, G., Castella, V., Stojanov, M., Baud, D., Vulliemoz, N., Ammerdorffer, A., Gyger, J., Greub, G., Castella, V., and Stojanov, M.
- Abstract
STUDY QUESTION What is the impact of Waddlia chondrophila, an emerging Chlamydia-related bacterium associated with miscarriage, on human spermatozoa? SUMMARY ANSWER W. chondrophila had a negative impact on human spermatozoa (decrease in viability and mitochondrial membrane potential) and was not entirely removed from infected samples by density gradient centrifugation. WHAT IS KNOWN ALREADY Bacterial infection or colonization might have a deleterious effect on male fertility. Waddlia chondrophila was previously associated with miscarriage, but its impact on male reproductive function has never been studied. STUDY DESIGN SIZE, DURATION An in vitro model of human spermatozoa infection was used to assess the effects of W. chondrophila infection. Controls included Chlamydia trachomatis serovar D and latex beads with similar size to bacteria. PARTICIPANTS/MATERIALS, SETTING, METHODS Purified motile spermatozoa were infected with W. chondrophila (multiplicity of infection of 1). Immunohistochemistry combined with confocal microscopy was used to evaluate how bacteria interact with spermatozoa. The impact on physiology was assessed by monitoring cell viability, mitochondrial membrane potential and DNA fragmentation. MAIN RESULTS AND THE ROLE OF CHANCE Using super-resolution confocal microscopy, bacteria were localized on spermatozoa surface, as well as inside the cytoplasm. Compared to controls, W. chondrophila caused a 20% increase in mortality over 72 h of incubation (P < 0.05). Moreover, higher bacterial loads significantly reduced mitochondrial membrane potential. Bacteria present on spermatozoa surface were able to further infect a cell-monolayer, indicating that sperm might vector bacteria during sexual intercourse. LIMITATIONS REASONS FOR CAUTION The main limitation of the study is the use of an in vitro model of infection, which might be too simplistic compared to an actual infection. An animal model of infection should be developed to better evaluate the i
- Published
- 2021
8. Forensic identification of urine samples: a comparison between nuclear and mitochondrial DNA markers
- Author
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Castella, V., Dimo-Simonin, N., Brandt-Casadevall, C., Robinson, N., Saugy, M., Taroni, F., Mangin, P., Castella, V., Dimo-Simonin, N., Brandt-Casadevall, C., Robinson, N., Saugy, M., Taroni, F., and Mangin, P.
- Abstract
Urine samples from 20 male volunteers of European Caucasian origin were stored at 4°C over a 4-month period in order to compare the identification potential of nuclear DNA (nDNA) and mitochondrial DNA (mtDNA) markers. The amount of nDNA recovered from urines dramatically declined over time. Consequently, nDNA likelihood ratios (LRs) greater than 1,000 were obtained for 100, 70 and 55% of the urines analysed after 6, 60 and 120 days, respectively. For the mtDNA, HVI and HVII sequences were obtained for all samples tested, whatever the period considered. Nevertheless, the highest mtDNA LR of 435 was relatively low compared to its nDNA equivalent. Indeed, LRs obtained with only three nDNA loci could easily exceed this value and are quite easier to obtain. Overall, the joint use of nDNA and mtDNA markers enabled the 20 urine samples to be identified, even after the 4-month period
- Published
- 2018
9. Waddlia chondrophila, a Chlamydia-related bacterium, has a negative impact on human spermatozoa
- Author
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Baud, D, primary, Vulliemoz, N, additional, Ammerdorffer, A, additional, Gyger, J, additional, Greub, G, additional, Castella, V, additional, and Stojanov, M, additional
- Published
- 2017
- Full Text
- View/download PDF
10. Driver or passenger? Use of a Bayesian network for the evaluation of DNA results in a fatal car accident.
- Author
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Samie L, Champod C, Hicks T, Delemont S, and Castella V
- Subjects
- Humans, Male, Automobile Driving, DNA genetics, DNA Fingerprinting, Forensic Genetics methods, Likelihood Functions, Accidents, Traffic, Bayes Theorem
- Abstract
This article presents a case where the issue was to determine who was the driver and who was the passenger at the time of a fatal car accident involving two persons, one of whom died in the accident. The presence of the two persons in the car was not contested, only the mechanisms that led to the deposition of the DNA (i.e., the activities) were. To our knowledge, few cases are evaluated considering the alleged activities. The reasons for this include the lack of knowledge, and data, as well as the difficulties encountered for the formulation of conclusions. In this case report, we present the architecture of the Bayesian Network (BN) used to evaluate the DNA results of the traces recovered from the steering wheel, driver's and passenger's airbags. The following propositions were considered: "The person of interest (POI) was driving the car and the alternative person (AP) was the passenger at the time of the accident" or vice versa. We discuss the assumptions that were made and how data from the literature was used to parametrize into the BN. A likelihood ratio of the order of 90 was finally assigned. The statement proposed to the mandating authority indicated that, given the information that was made available to us, our observations were of the order of 90 times more probable if the POI was driving the car at the time of the accident rather than if the AP was. A sensitivity analysis was performed (5000 simulations): this shows that our likelihood ratio is robust., Competing Interests: Declaration of Competing Interest No conflict of interest., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2025
- Full Text
- View/download PDF
11. Searching national DNA databases with complex DNA profiles: An empirical study using probabilistic genotyping.
- Author
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Nozownik S, Hicks T, Basset P, and Castella V
- Abstract
In most National DNA databases (NDNADB), only single source DNA profiles, and sometimes two-person DNA mixtures, can be searched provided a minimum number of loci (or alleles) is available. DNA profiles that do not meet these criteria (about 14 % of the traces analyzed in Western Switzerland) can be compared locally with candidates upon request from police services, used for one-off search, or remain unused. With the advent of probabilistic genotyping (PG), such complex DNA profiles can be compared to those stored in NDNADB based on likelihood ratios (LRs). In this pilot study, traces of known contributors and casework DNA profiles were used to evaluate the performance of the DBLR™ "Search database" tool in conjunction with the Swiss NDNADB. First, 40 DNA mixtures (2-5 contributors) from 15 volunteers were prepared in the wet laboratory. They were deconvoluted with STRmix™ and compared to a database containing the DNA profiles of these 15 volunteers, along with 174,493 person DNA profiles from the Swiss NDNADB (ground-truth experiments). Using LR thresholds of 10
3 and 106 , sensitivity and specificity were respectively 90.0 %/57.1 % and 99.9 %/100.0 %. For the lower LR threshold, this resulted in 52 adventitious associations out of more than 24 million pairwise comparisons. Second, 160 DNA mixture profiles from casework (2-4 contributors) that had previously been locally compared were searched with DBLR™ using the same conditions as for phase 1. With the 103 LR threshold, 380 associations were retrieved: 194 of these corresponded to expected associations, as they were previously made through the local comparisons with known persons, and 186 were new. With the 106 LR threshold, 199 associations were recovered of which 180 were expected and 19 new. This demonstrates that even with complex DNA profiles (up to 4 contributors) all expected associations were retrieved with a limited number of candidates per trace. Database searches of complex DNA mixtures allow for the generation of leads early in an investigation for DNA profiles that might otherwise remain underutilized. Next steps for the possible integration of DBLR™ or similar software within an operational context will require discussions on legal, financial, and technical aspects among stakeholders., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)- Published
- 2024
- Full Text
- View/download PDF
12. Value of DNA mixture-to-mixture comparisons within an operational context.
- Author
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Basset P, Toulemont L, Hicks T, and Castella V
- Subjects
- Humans, Switzerland, Likelihood Functions, Genotype, Police, DNA Fingerprinting, Microsatellite Repeats, DNA genetics, Software, Databases, Nucleic Acid
- Abstract
Since 1995, national forensic DNA databases have used a maximum number of contributors, and a minimum number of loci to reduce the risk of providing false leads. DNA profiles of biological traces that do not meet these criteria cannot be loaded into these databases. In 2023, about 10 % of more than 15,000 trace DNA profiles analyzed in western Switzerland were not compared at the national level, even though they were considered to be interpretable, mainly because they contained the DNA from more than two persons. In this situation, police services can request local comparisons with DNA profiles of known persons and/or with other traces, but this occurs in only a small proportion of cases, so that DNA mixtures are rarely used to help detect potential series. The development of probabilistic genotyping software and its associated tools have made possible the efficient performance of this type of comparison, which is based on likelihood ratios (LR) rather than on the number of shared alleles. To highlight potential common contributors for investigation and intelligence purposes, the present study used the mixture-to-mixture tool of the software STRmix v2.7 to compare 235 DNA profiles that cannot be searched the Swiss DNA database. These DNA profiles originated from traces collected by six different police services in 2021 and 2022. Traces were selected by the police based on information that indicated that they were from potential series. Associations between profiles were compared with expected investigative associations to define the value of this approach. Among the 27,495 pairwise comparisons of DNA profiles, 88 pairs (0.3 %) showed at least one potential common contributor when using a LR threshold of 1000. Of these 88 pairs, 60 (68.2 %) were qualified by the police services as "expected" (60/88), 22 (25.0 %) as "possible", and six (6.8 %) as "unexpected". Although it is important to consider the limits of this approach (e.g., adventitious or missed associations, cost/benefit evaluation, integration of DNA mixture comparison in the process), these findings indicate that non CODIS loadable DNA mixtures could provide police agencies with information concerning potential series at both the local and national level., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2024
- Full Text
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13. Characterization of DNA concentration in urine and dried blood samples to detect the c.577 deletion within the EPO gene.
- Author
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Leuenberger N, Jan N, Kuuranne T, and Castella V
- Subjects
- Humans, Male, Female, Sequence Analysis, DNA methods, Sequence Deletion, Erythropoietin urine, Erythropoietin genetics, Erythropoietin blood, Doping in Sports, DNA urine, DNA genetics, DNA blood, Dried Blood Spot Testing methods, Substance Abuse Detection methods
- Abstract
The EPO gene variant, c.577del (VAR-EPO), was discovered in the Chinese population in 2021. The mutated protein is naturally present in urine from individuals heterozygous for the variant. Electrophoresis methods currently applied in anti-doping laboratories produce a pattern in samples from individuals carrying VAR-EPO that cannot be unambiguously distinguished from individuals who received recombinant EPO doses. Consequently, the analysis of blood samples is obligatory to facilitate interpretation of suspicious findings from urine samples. However, this complicates the process and delays the reporting. Objective of this study was to develop EPO c.577del detection in urine and dried blood samples (DBS) in order to facilitate and accelerate EPO results management. Moreover, estimation of the success rate of sequencing regarding concentration of DNA in urine and DBS was evaluated. Conclusive results regarding Sanger sequencing were obtained for all samples with DNA concentrations above 0.024 ng/μL DNA in 80% of urines samples from volunteers. The potential success of DNA sequencing rate in athletes' urines was investigated. A total of 191 urine samples were considered. DNA concentration exceeding 0.024 ng/μL was detected in 85% of the samples. Interestingly, in-competition samples had a significantly higher DNA concentration than out-of-competition male urine samples (0.330 vs. 0.084 ng/μL). Moreover, conclusive EPO sequences were obtained for 100% of DBS (cellulose and polymer matrices). In conclusion, method for detection of EPO gene variant was developed in urine and DBS. Characterization of DNA concentration was performed in order to evaluate the probability of success of sequencing EPO gene in anti-doping field., (© 2024 John Wiley & Sons Ltd.)
- Published
- 2024
- Full Text
- View/download PDF
14. Comparison between MACSprep™ forensic sperm microbead kit and Erase Sperm Isolation kit for the enrichment of sperm fractions recovered from sexual assault samples.
- Author
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Grosjean F, Favre M, and Castella V
- Subjects
- Humans, Male, Female, Microspheres, Spermatozoa, DNA, DNA Fingerprinting, Azoospermia, Sex Offenses
- Abstract
Sexual assault samples often contain mixtures of cells coming from at least two donors. Ideally, one would need to separate the cells into two cellular fractions: one consisting of the alleged aggressor's spermatozoa (the sperm fraction) and the other containing the victim's epithelial cells (the non-sperm fraction). This separation increases the probability of obtaining the alleged offender's autosomal DNA profile. However, spermatozoa are often collected along with an excess of biological material originating from the victim, and with unfavorable male:female biological material ratios, the absence of separation could result in the PCR amplification of the victim's DNA profile only. Several approaches are available to enrich/purify the spermatozoa present on sexual assault samples. In this paper, we compare a new method, the MACSprep™ Forensic Sperm MicroBead Kit (MACSprep, based on microbeads conjugated with antibodies bound to spermatozoa and their retention within a magnetic column) with the Erase Sperm Isolation Kit (Erase, a standard differential lysis separation procedure combined with a specific removal of free DNA) routinely used in our lab. The performance of both kits was tested using sets of vaginal and buccal swabs loaded with different dilutions of sperm, or azoospermic semen, representing a total of 120 independent samples. For the samples containing undiluted sperm, an average recovery of 58% was observed for the MACSprep's sperm fractions and 43% for Erase's. Significantly better recovery of azoospermic semen was observed in MACSprep's non-sperm fractions (~ 85%) compared to Erase (~ 28%). Erase performed significantly better than MACSprep in terms of recovery for diluted sperm samples (1:10 to 1:800 sperm dilutions) in the presence of vaginal cells, while the purities of the achieved sperm fractions were in favor of MACSprep for the highest sperm dilutions tested. Similar trends were observed with buccal swabs loaded with 1:200 sperm dilutions. Increased sperm dilutions on vaginal swabs resulted in higher variability in the male material recovered, whatever the separation method used. Both methods were easy to perform and resulted in male DNA extracts ready to use in less than 2 h. Both kits showed their specificities in terms of recovery efficiency and purity of the sperm fractions. Ideally, additional experiments should be performed in different laboratories, using workflow and chemistries different than ours, to better define the peculiarities observed with MACSprep for high dilutions. Improving the recovery of MACSprep for diluted samples, in addition to its better purity observed in the experiments performed, could make it a method of choice for laboratory workflow, despite MACSprep's current price per sample being about twice the price of Erase's., (© 2022. The Author(s).)
- Published
- 2023
- Full Text
- View/download PDF
15. A simplified protocol for the detection of blood, saliva, and semen from a single biological trace using immunochromatographic tests.
- Author
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Basset P, Blandin P, Grini A, Delemont S, Samie L, and Castella V
- Subjects
- DNA analysis, DNA Fingerprinting methods, Humans, Male, Saliva chemistry, Body Fluids, Semen chemistry
- Abstract
The detection of body fluids (e.g., blood, saliva or semen) provides information that is important both for the investigation and for the choice of the analytical protocols. Because of their sensitivity, specificity, as well as their simplicity of use, immunochromatographic tests are widely applied. These tests target different body fluids and generally require specific buffer solutions. If one needs to investigate whether the material is of a specific nature (e.g., blood), this is fine. However, if the material can also contain other material (e.g., saliva or semen) then the use of different tests can be problematic. Indeed, if the different tests require different buffers, it will not be possible to perform all tests on the exact same specimen.In this study, we assess the use of the RSID™-universal buffer to perform three immunochromatographic tests (HEXAGON OBTI, RSID-saliva, and PSA Semiquant) as well as spermatozoa detection. We use the same eluate for the detection of all three body fluids. The proposed protocol provides similar results to those obtained when each test is conducted independently. Furthermore, it does not affect the quality of the DNA profiles. The main advantage of this protocol is that the results of the presumptive test(s) and of the DNA analyses are representative of the exact same specimen., (© 2022. The Author(s).)
- Published
- 2022
- Full Text
- View/download PDF
16. A Logical Framework for Forensic DNA Interpretation.
- Author
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Hicks T, Buckleton J, Castella V, Evett I, and Jackson G
- Subjects
- DNA genetics
- Abstract
The forensic community has devoted much effort over the last decades to the development of a logical framework for forensic interpretation, which is essential for the safe administration of justice. We review the research and guidelines that have been published and provide examples of how to implement them in casework. After a discussion on uncertainty in the criminal trial and the roles that the DNA scientist may take, we present the principles of interpretation for evaluative reporting. We show how their application helps to avoid a common fallacy and present strategies that DNA scientists can apply so that they do not transpose the conditional. We then discuss the hierarchy of propositions and explain why it is considered a fundamental concept for the evaluation of biological results and the differences between assessing results given propositions that are at the source level or the activity level. We show the importance of pre-assessment, especially when the questions relate to the alleged activities, and when transfer and persistence need to be considered by the scientists to guide the court. We conclude with a discussion on statement writing and testimony. This provides guidance on how DNA scientists can report in a balanced, transparent, and logical way.
- Published
- 2022
- Full Text
- View/download PDF
17. Use of Bayesian Networks for the investigation of the nature of biological material in casework.
- Author
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Samie L, Champod C, Delémont S, Basset P, Hicks T, and Castella V
- Subjects
- Bayes Theorem, Forensic Medicine, Saliva
- Abstract
Chemical and staining methods, immunochromatography, spectroscopy, RNA expression or methylation patterns, do not allow to determine the nature of the biological material with certainty. However, to our knowledge, there are few forensic scientists that assess the value of such test results using a probabilistic approach. This is surprising as it would allow account for false positives and false negatives and avoid misleading conclusions. In this paper, we developed three Bayesian Networks (BNs) to assess the presence of blood, saliva and sperm in the recovered material and combine potentially contradictory observations. The approach was successfully tested using 188 traces from proficiency tests. We have implemented an online user-friendly application (https://forensic-genetic.shinyapps.io/BodyFluidsApp/) that allows forensic scientists to assess the value of their results without having to build Bayesian Networks themselves. They can also input their own data, use the application to identify a potential lack of knowledge and report their conclusions regarding the presence of sperm, blood or/and saliva considering uncertainty., Competing Interests: Conflict of Interest No conflict of interest., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2022
- Full Text
- View/download PDF
18. The Lady from Basel's Barfüsserkirche - Molecular confirmation of the Mummy's identity through mitochondrial DNA of living relatives spanning 22 generations.
- Author
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Wurst C, Maixner F, Castella V, Cipollini G, Hotz G, and Zink A
- Subjects
- DNA, Mitochondrial genetics, Haplotypes, Humans, Sequence Analysis, DNA, Genome, Mitochondrial, Mummies
- Abstract
The identity of the mummified Lady from the Barfüsser Church in Basel, Switzerland has been unsolved for decades, despite the prominent location of the burial place in front of the choir screen. A recent multidisciplinary research approach came up with a possible candidate, Anna Catharina Bischoff who died in Basel in 1787 with an age of 69 years (1719-1787). To verify the identity of the mummy, genealogists of the Citizen Science Basel discovered three living individuals of the maternal lineage of two different family branches, separated from Anna Catharina Bischoff by up to 22 generations. In this study we compare the ancient mitochondrial DNA of the mummy recovered from a premolar to the mitochondrial DNA of these three candidates. Initially the mitochondrial hypervariable regions I and II of the living individuals were screened using the Sanger sequencing method. This was followed by a mitochondrial capture approach and next generation sequencing to enrich for the whole mitochondrial genome of the mummy and one living person. A full mitochondrial genome has been recovered of both individuals sharing an identical haplotype. The sequence was assigned to the mitochondrial haplogroup U5a1+!16192 including two private mutations 10006G and 16293C. Only by using an interdisciplinary approach combining ancient DNA analysis and genealogy a maternal lineage of a non-noble family spanning 22 generations could be confirmed., (Copyright © 2021 Elsevier B.V. All rights reserved.)
- Published
- 2022
- Full Text
- View/download PDF
19. A Swiss collaborative exercise for Disaster Victim Identification (DVI): Covering situations with different levels of complexity.
- Author
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Gehrig C, Delémont S, Comte J, Hicks T, Basset P, Grosjean F, Dion D, Cossu C, Bottinelli M, Hecht M, Sulzer A, Voegeli P, and Castella V
- Subjects
- Adult, Child, Humans, Switzerland, DNA Fingerprinting, Disaster Victims, Forensic Anthropology, Forensic Genetics, Pedigree, Simulation Training, Software
- Abstract
The identification of victims of a disaster (DVI) requires the collaboration of different specialists. Within a DVI context, DNA analyses often play an important role. Consequently, forensic genetic laboratories should be prepared to cope with DVI situations, as this can involve large-scale DNA profile comparisons. Six forensic genetic laboratories from Switzerland participated in an exercise where supposedly a plane had crashed. The goal of the exercise was to monitor participants use of dedicated software with ground truth cases and to make them aware of the existence of particular situations that may occur in real cases. For assigning the value of the comparison of the DNA profiles, all participating laboratories used the DVI module of Familias v3.2.
1 In addition, one of the 6 laboratories used the Pedigree Searcher from CODIS v7.0. The data (AmpFlSTR® NGM SElect™ profiles) were generated to challenge the participating laboratories: cases with first, second degree biological parents, mutation events, as well as non-paternity cases were included. This study shows that the majority of the participants used the software in an appropriate way. However, a few misleading conclusions were detected for the most challenging situations. These errors belonged to one of the following categories: false pedigree, false association using the higher LR, misleading contextual information (false paternity) and not clustering family members. Specific recommendations are provided in order to reduce misuse of the software and the risk of misinterpretations by using all the relevant information., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)- Published
- 2021
- Full Text
- View/download PDF
20. Touch DNA collection - Performance of four different swabs.
- Author
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Comte J, Baechler S, Gervaix J, Lock E, Milon MP, Delémont O, and Castella V
- Subjects
- DNA Fingerprinting, Humans, Preservation, Biological, Real-Time Polymerase Chain Reaction, DNA analysis, Specimen Handling instrumentation, Touch
- Abstract
A collaborative study conducted by three police forensic units, a DNA laboratory, and a forensic academic institute was undertaken in order to compare the performance of four different swabs in controlled and quasi-operational conditions. For this purpose, a reference swab (Prionics cardboard evidence collection kit) currently used within the police forensic units and 3 challenger swabs (COPAN 4N6FLOQSwabs™ (Genetics variety), Puritan FAB-MINI-AP and Sarstedt Forensic Swab) were used for collecting DNA traces from previously used items (referred as "touch DNA" in this article) including on 60 collars, 60 screwdrivers and 60 steering wheels obtained from volunteers. For each comparison, the surface considered was divided into two equal components; one was sampled with the reference swab and the other with one of the three challenger swabs. This lead to a total of 360 samples. Conclusions were consistent within the four operational partners. From a practical point of view, the COPAN 4N6FLOQSwabs™ (Genetics variety) was judged the most convenient to use. Furthermore, it allowed the recovery of significantly more DNA from collars (0.65 vs 0.13 ng/μL) and steering wheels (2.82 vs 1.77 ng/μL), and a similar amount of DNA from screwdrivers (0.032 vs 0.026 ng/μL) compared with the Prionics reference swab. The two other challenger swabs provided results that were not significantly different from the reference swab, except for the Puritan swab, whose performance was significantly lower for steering wheels (0.37 vs 0.58 ng/μL). As part of a conservation study, 50 μL of a blood dilution (1/4 with PBS) was deposited on a total of 105 COPAN (Genetics and Crime Scene varieties), Prionics and Sarstedt swabs. They were stored within a cupboard at room temperature. The integrity of the recovered DNA was evaluated with NGM SElect™ DNA profiles after different time-spans ranging from 1 day to 12 months by comparing the height difference of the peaks occurring at the shortest and longest loci, respectively. DNA seemed to remain stable, except when using the COPAN 4N6FLOQSwabs™ treated with an antimicrobial agent (Crime scene variety), which resulted in significant DNA degradation. Following these tests, the COPAN 4N6FLOQSwabs™ (Genetics variety), a model with a desiccant, was selected for further testing in fully operational conditions., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2019
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- View/download PDF
21. The efficiency of DNA extraction kit and the efficiency of recovery techniques to release DNA using flow cytometry.
- Author
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Samie L, Champod C, Glutz V, Garcia M, Castella V, and Taroni F
- Subjects
- Genetic Techniques instrumentation, Humans, Keratinocytes, Laboratories, Polymerase Chain Reaction, DNA isolation & purification, Efficiency, Flow Cytometry, Reagent Kits, Diagnostic
- Published
- 2019
- Full Text
- View/download PDF
22. Sperm Microbiota and Its Impact on Semen Parameters.
- Author
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Baud D, Pattaroni C, Vulliemoz N, Castella V, Marsland BJ, and Stojanov M
- Abstract
Compared to its female counterpart, the microbiota of the male genital tract has not been studied extensively. With this study, we aimed to evaluate the bacterial composition of seminal fluid and its impact on sperm parameters. We hypothesized that a dysbiotic microbiota composition may have an influence on sperm quality. Semen samples of 26 men with normal spermiogram and 68 men with at least one abnormal spermiogram parameter were included in the study. Samples were stratified based on total sperm count, spermatozoa concentration, progressive motility, total motility and spermatozoa morphology. Microbiota profiling was performed using 16S rRNA gene amplicons sequencing and total bacterial load was determined using a panbacterial quantitative PCR. Semen samples broadly clustered into three microbiota profiles: Prevotella -enriched, Lactobacillus -enriched, and polymicrobial. Prevotella -enriched samples had the highest bacterial load ( p < 0.05). Network analysis identified three main co-occurrence modules, among which two contained bacteria commonly found in the vaginal flora. Genera from the same module displayed similar oxygen requirements, arguing for the presence of different ecological niches for bacteria that colonize semen through the passage. Contrary to our hypothesis, shifts in overall microbiota composition (beta-diversity) did not correlate with spermiogram parameters. Similarly, we did not find any difference in microbial richness or diversity (alpha-diversity). Differential abundance testing, however, revealed three specific genera that were significantly enriched or depleted in some of the sperm quality groups ( p < 0.05). Prevotella relative abundance was increased in samples with defective sperm motility while Staphylococcus was increased in the corresponding control group. In addition, we observed an increased relative abundance of Lactobacillus in samples with normal sperm morphology. Our study indicates that overall bacterial content of sperm might not play a major role in male infertility. Although no major shifts in microbiota composition or diversity were found, the differential abundance of specific bacterial genera in the sperm suggests that a small subset of microbes might impact the spermatozoal physiology during sperm transition, more specifically motility and morphology. Further studies are required to challenge this finding and develop potential strategies to induce the formation of a healthy seminal microbiota.
- Published
- 2019
- Full Text
- View/download PDF
23. Positive impact of DNA contamination minimization procedures taken within the laboratory.
- Author
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Basset P and Castella V
- Subjects
- Humans, Quality Control, Switzerland, DNA Contamination, Laboratories, Specimen Handling standards
- Abstract
DNA contamination incidents are one of the most frequent sources of error in forensic genetics and can have serious consequences. It is therefore essential to take measures to prevent these events and to monitor the real impact of contamination minimization procedures. In this study, we review and compare the number of contamination events detected on trace samples analyzed by the Forensic Genetic Unit (FGU) of the University Center of Legal Medicine in Switzerland before and after the implementation of new contamination minimization procedures. Interestingly, the number of contamination events by laboratory staff was significantly reduced by more than 70% after the implementation of the procedures. However, no significant change was observed for contamination events by police collaborators. This difference is likely to be explained by the differential impact of procedures taken in the laboratory and on crime scene. It suggests that the reduction observed for laboratory contamination incidents is due to the new procedures taken. In conclusion, our study highlights that taking appropriate measures is efficient and can reduce the number of contamination incidents. However, it is important that such contamination minimization procedures be implemented all along the chain of analysis of a stain (i.e. from crime scene to the laboratory)., (Copyright © 2018 Elsevier B.V. All rights reserved.)
- Published
- 2019
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24. Lessons from a study of DNA contaminations from police services and forensic laboratories in Switzerland.
- Author
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Basset P and Castella V
- Subjects
- Humans, Retrospective Studies, Surveys and Questionnaires, Switzerland, DNA Contamination, Laboratories statistics & numerical data, Police statistics & numerical data
- Abstract
In Switzerland, the DNA profiles of police officers collecting crime scene traces as well as forensic genetic laboratories employees are stored in the staff index of the national DNA database to detect potential contaminations. Our study aimed at making a national inventory of contaminations to better understand their origin and to make recommendations in order to decrease their occurrence. For this purpose, a retrospective questionnaire was sent to both police services and forensic genetic laboratories for each case where there was a contamination. Between 2011 and 2015, a total of 709 contaminations were detected. This represents a mean of 11.5 (9.6-13.4) contaminations per year per 1'000 profiles sent to the Swiss DNA database. Feedbacks were obtained from the police, the laboratory or both for 552/709 (78%) of the contaminations. Approximately 86% of these contaminations originated from police officers whereas only 11% were from genetic laboratories employees and 3% were associated to other sources (e.g. positive controls, stain-stain contaminations). Interestingly, a direct contact between the stain and the contaminant person occurred in only 51% of the laboratory contaminations whereas this number increased to 91% for police collaborators. The high level of indirect DNA transfer in laboratories might be explained by the presence of "DNA reservoirs" suggesting that cleaning procedures should be improved. At the police level, most contaminations originated from the person who collected the trace and likely occurred directly at the crime scene. Improving sampling practices could be beneficial to reduce these contaminations., (Copyright © 2017 Elsevier B.V. All rights reserved.)
- Published
- 2018
- Full Text
- View/download PDF
25. Application of DIP-STRs to sexual/physical assault investigations: Eight case reports.
- Author
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Oldoni F, Castella V, and Hall D
- Subjects
- Chromosomes, Human, Y, Electrophoresis, Capillary, Female, Humans, Male, Polymerase Chain Reaction, DNA Fingerprinting methods, INDEL Mutation, Microsatellite Repeats, Physical Abuse, Polymorphism, Genetic, Sex Offenses
- Abstract
DIP-STRs are compound markers formed by a deletion/insertion polymorphism linked to a microsatellite. They enable the deconvolution of unbalanced DNA mixtures from two individuals, up to 1000 fold excess of one contributor. In practice, this novel tool allows to test for the presence of a DNA of interest in traces appearing not useful because of the masking effect of the major DNA contributor. Thus far two sets of DIP-STRs have been published: the first set was described as proof-of-principle, while the second set was specifically developed for forensic applications. Here, we report on the first use of these markers in casework to show advantages and limitations in real examples. Traces, suggestive of containing unbalanced DNA mixtures (beyond standard STR mixture resolution), were selected from eight cases submitted to the Forensic Genetics Unit of the University Center of Legal Medicine of Lausanne-Geneva. Using 18 validated DIP-STRs, two to ten markers were selected for each case. A minor DNA contributor - undetected using conventional STRs - was detected for the trace samples of six cases. DIP-STR results contributed to each case, either by complementing Y-STRs results or by producing novel investigative leads. This was especially true with same sex unbalanced DNA mixtures, female minor/male major unbalanced DNA mixtures or when the source of the DNA mixture was said to come either from the suspect and the female complainant or from his brother and the female complainant. Interestingly, these markers were found to be more sensitive and specific than previously known. Positive results were obtained at 16,000-fold excess of major DNA using few picograms of input DNA, as well as from traces collected several months after the alleged offence. Likelihood ratios assigned to measure the strength of DIP-STRs' DNA evidence were modest (10), when accounted by only two DIP-STRs, and high (10
6 ) when determined by six markers. In some cases the detection of extra alleles from additional minor DNA contributors or because of extremely unbalanced DNA ratios, limited the interpretation of the results. In conclusion, the DIP-STRs often provide additional value to the analysis of traces that cannot be exploited by the use of standard methods., (Copyright © 2017 Elsevier B.V. All rights reserved.)- Published
- 2017
- Full Text
- View/download PDF
26. Sensitive DIP-STR markers for the analysis of unbalanced mixtures from "touch" DNA samples.
- Author
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Oldoni F, Castella V, Grosjean F, and Hall D
- Subjects
- Chromosomes, Human, Y, DNA Fingerprinting, Genetic Markers, Humans, Male, Mutagenesis, Insertional, Polymerase Chain Reaction, DNA genetics, Genotyping Techniques, Microsatellite Repeats, Polymorphism, Single Nucleotide, Touch
- Abstract
Casework samples collected for forensic DNA analysis can produce genomic mixtures in which the DNA of the alleged offender is masked by high quantities of DNA coming from the victim. DIP-STRs are novel genetic markers specifically developed to enable the target analysis of a DNA of interest in the presence of exceeding quantities of a second DNA (up to 1000-fold). The genotyping system, which is based on allele-specific amplifications of haplotypes formed by a deletion/insertion polymorphism (DIP) and a short tandem repeat (STR), combines the capacity of targeting the DNA of an individual with a strong identification power. Finally, DIP-STRs are autosomal markers therefore they can be applied to any combination of major and minor DNA. In this study we aimed to assess the ability of DIP-STRs to detect the minor contributor on challenging "touch" DNA samples simulated with representative crime-associated substrates and to compare their performance to commonly used male-specific markers (Y-STRs). As part of a comprehensive study on the relative DNA contribution of two persons handling the same object, we selected 71 unbalanced contact traces of which 14 comprised a male minor DNA contributor mixed to a female major DNA contributor. Using a set of six DIP-STRs, one to four markers were found to be informative for the minor DNA detection across traces. When compared to Y-STRs (14 traces), the DIP-STRs showed similar sensitivity in detecting the minor DNA across substrate materials with a similar occurrence of allele drop-out. Conversely, because of the sex combination of the two users of the object, 57 remaining traces could only be investigated by DIP-STRs. Of these, 30 minor DNA contributors could be detected by all informative markers while 12 traces showed events of allele drop-out. Finally, 15 traces showed no amplification of the minor DNA. These last 15 samples were mostly characterized by a combination of short handling time of the object, low DNA recovery and/or one single informative DIP-STR. In conclusion, the DIP-STRs represent alternative markers to help solving unbalanced two-source DNA mixtures, and also those produced from contact stains. These markers, in addition to a novel set of 10 DIP-STRs specifically developed according to forensic technical standards, will offer a valuable tool complementary to Y-STR markers., (Copyright © 2017 Elsevier B.V. All rights reserved.)
- Published
- 2017
- Full Text
- View/download PDF
27. Stabbing simulations and DNA transfer.
- Author
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Samie L, Hicks T, Castella V, and Taroni F
- Subjects
- Alleles, Computer Simulation, DNA genetics, DNA Fingerprinting statistics & numerical data, Humans, Probability, DNA isolation & purification, DNA Fingerprinting methods, Forensic Genetics methods, Wounds, Stab genetics
- Abstract
Technical developments have made it possible to analyze very low amounts of DNA. This has many advantages, but the drawback of this technological progress is that interpretation of the results becomes increasingly complex: the number of mixed DNA profiles increased relatively to single source DNA profiles and stochastic effects in the DNA profile, such as drop-in and drop-out, are more frequently observed. Moreover, the relevance of low template DNA material regarding the activities alleged is not as straightforward as it was a few years ago, when for example large quantities of blood were recovered. The possibility of secondary and tertiary transfer is now becoming an issue. The purpose of this research is twofold: first, to study the transfer of DNA from the handler and secondly, to observe if handlers would transfer DNA from persons closely connected to them. We chose to mimic cases where the offender would attack a person with a knife. As a first approach, we envisaged that the defense would not give an alternative explanation for the origin of the DNA. In our transfer experiments (4 donors, 16 experiments each, 64 traces), 3% of the traces were single DNA profiles. Most of the time, the DNA profile of the person handling the knife was present as the major profile: in 83% of the traces the major contributor profile corresponded to the stabber's DNA profile (in single stains and mixtures). Mixture with no clear major/minor fraction (12%) were observed. 5% of the traces were considered of insufficient quality (more than 3 contributors, presence of a few minor peaks). In that case, we considered that the stabber's DNA was absent. In our experiments, no traces allowed excluding the stabber, however it must be noted that precautions were taken to minimize background DNA as knives were cleaned before the experiments. DNA profiles of the stabber's colleagues were not observed. We hope that this study will allow for a better understanding of the transfer mechanism and of how to assess and describe results given activity level propositions. In this preliminary research, we have focused on the transfer of DNA on the hand of the person. Besides, more research is needed to assign the probability of the results given an alternative activity proposed by the defense, for instance when the source of the DNA is not contested, but that the activities are., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)
- Published
- 2016
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28. (210)Po poisoning as possible cause of death: forensic investigations and toxicological analysis of the remains of Yasser Arafat.
- Author
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Froidevaux P, Bochud F, Baechler S, Castella V, Augsburger M, Bailat C, Michaud K, Straub M, Pecchia M, Jenk TM, Uldin T, and Mangin P
- Subjects
- Bayes Theorem, Cause of Death, France, Humans, Lead Radioisotopes analysis, Lead Radioisotopes poisoning, Radioisotopes, Famous Persons, Forensic Toxicology methods, Polonium poisoning
- Abstract
The late president of the Palestinian Authority, Yasser Arafat, died in November 2004 in Percy Hospital, one month after having experienced a sudden onset of symptoms that included severe nausea, vomiting, diarrhoea and abdominal pain and which were followed by multiple organ failure. In spite of numerous investigations performed in France, the pathophysiological mechanisms at the origin of the symptoms could not be identified. In 2011, we found abnormal levels of polonium-210 ((210)Po) in some of Arafat's belongings that were worn during his final hospital stay and which were stained with biological fluids. This finding led to the exhumation of Arafat's remains in 2012. Significantly higher (up to 20 times) activities of (210)Po and lead-210 ((210)Pb) were found in the ribs, iliac crest and sternum specimens compared to reference samples from the literature (p-value <1%). In all specimens from the tomb, (210)Po activity was supported by a similar activity of (210)Pb. Biokinetic calculations demonstrated that a (210)Pb impurity, as identified in a commercial source of 3MBq of (210)Po, may be responsible for the activities measured in Arafat's belongings and remains 8 years after his death. The absence of myelosuppression and hair loss in Mr Arafat's case compared to Mr Litvinenko's, the only known case of malicious poisoning with (210)Po, could be explained by differences in the time delivery-scheme of intake. In conclusion, statistical Bayesian analysis combining all the evidence gathered in our forensic expert report moderately supports the proposition that Mr Arafat was poisoned by (210)Po., (Copyright © 2015 The Authors. Published by Elsevier Ireland Ltd.. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
29. A novel set of DIP-STR markers for improved analysis of challenging DNA mixtures.
- Author
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Oldoni F, Castella V, and Hall D
- Subjects
- Gene Frequency, Haplotypes, Humans, Limit of Detection, Microsatellite Repeats genetics, Polymorphism, Genetic, DNA genetics, Forensic Genetics, Genetic Markers
- Abstract
Currently available molecular biology tools allow forensic scientists to characterize DNA evidence found at crime scenes for a large variety of samples, including those of limited quantity and quality, and achieve high levels of individualization. Yet, standard forensic markers provide limited or no results when applied to mixed DNA samples where the contributors are present in very different proportions (unbalanced DNA mixtures). This becomes an issue mostly for the analysis of trace samples collected on the victim or from touched objects. To this end, we recently proposed an innovative type of genetic marker, named DIP-STR that relies on pairing deletion/insertion polymorphisms (DIP) with standard short tandem repeats (STR). This novel compound marker allows detection of the minor DNA contributor in a DNA mixture of any gender and cellular origin with unprecedented resolution (beyond a DNA ratio of 1:1000). To provide a novel analytical tool useful in practice to common forensic laboratories, this article describes the first set of 10 DIP-STR markers selected according to forensic technical standards. The novel DIP-STR regions are short (between 146 and 271 bp), include only highly polymorphic tri-, tetra- and pentanucleotide tandem repeats and are located on different chromosomes or chromosomal arms to provide statistically independent results. This novel set of DIP-STR can target the amplification of 0.03-0.1 ng of DNA when mixed with a 1000-fold excess of major DNA. DIP-STR relative allele frequencies are estimated based on a survey of 103 Swiss individuals. Finally, this study provides an estimate of the occurrence of informative alleles and a calculation of the corresponding random match probability of the detected minor DIP-STR genotype assessed across 10,506 pairwise conceptual mixtures., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)
- Published
- 2015
- Full Text
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30. Catching allergy by a simple questionnaire.
- Author
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Galimberti M, Passalacqua G, Incorvaia C, Castella V, Costantino MT, Cucchi B, Gangemi S, Nardi G, Raviolo P, Rottoli P, Scichilone N, Sciolla N, Bettoncelli G, Landi M, Ridolo E, Buttafava S, Puccinelli P, Canonica GW, Fiocchi A, and Frati F
- Abstract
Background: Identifying allergic rhinitis requires allergy testing, but the first-line referral for rhinitis are usually primary care physicians (PCP), who are not familiar with such tests. The availability of easy and simple tests to be used by PCP to suggest allergy should be very useful., Methods: The Respiratory Allergy Prediction (RAP) test, based on 9 questions and previously validated by a panel of experts, was evaluated in this study., Results: An overall number of 401 patients (48.6% males, age range 14-62 years) with respiratory symptoms was included. Of them, 89 (22.2%) showed negative results to SPT, while 312 (77.8%) had at least one positive result to SPT. Cohen's kappa coefficient showed that all questions had an almost perfect excellent agreement between pre and post-test. The algorithm of decision-tree growth Chi-squared Automatic Interaction Detector showed that answering yes to the question 4 (Your nasal/ocular complains do usually start or worsen during the spring?), 6 (Did you ever had cough or shortness of breath, even during exercise?) and 8 (Do you use nasal sprays frequently?) gave a probability to have a positive SPT of 85%., Conclusions: These findings show that RAP test can be proposed as an useful tool to be used by physician other than allergists when evaluating patients with rhinitis, suggesting the need of allergy testing.
- Published
- 2015
- Full Text
- View/download PDF
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