Our research has developed a highly sensitive and simple assay to detect small amounts of animal and human biological material in less than 40 min. The handheld SaLux19 device developed at the Max Planck Institute of Experimental Medicine in Göttingen, Germany, was used to validate our concept. The proposed system uses isothermal amplification of DNA in a rapid assay format. Our results show that the assay can detect Sus scrofa nucleic acids with very high sensitivity and specificity. This detection system has potential for forensic scenarios.
- Publikační typ
- časopisecké články MeSH
Research on ancient and forensic DNA is related in many ways, and the two fields must deal with similar obstacles. Therefore, communication between these two communities has the potential to improve results in both research fields. Here, we present the insights gained in the ancient DNA community with regard to analyzing DNA from aged skeletal material and the potential use of the developed protocols in forensic work. We discuss the various steps, from choosing samples for DNA extraction to deciding between classical PCR amplification and massively parallel sequencing approaches. Based on the progress made in ancient DNA analyses combined with the requirements of forensic work, we suggest that there is substantial potential for incorporating ancient DNA approaches into forensic protocols, a process that has already begun to a considerable extent. However, taking full advantage of the experiences gained from ancient DNA work will require comparative studies by the forensic DNA community to tailor the methods developed for ancient samples to the specific needs of forensic studies and case work. If successful, in our view, the benefits for both communities would be considerable.
- MeSH
- DNA fingerprinting MeSH
- DNA * genetika MeSH
- lidé MeSH
- nekrotická degradace DNA MeSH
- senioři MeSH
- soudní genetika MeSH
- starobylá DNA * MeSH
- Check Tag
- lidé MeSH
- senioři MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Skeletal remains are among the most difficult types of samples encountered in forensic DNA casework and historical investigations due to prolonged exposure to environmental insults. DNA extracted from bone often is degraded, in low quantities, and contains co-purified inhibitors from the surrounding soil and/or burial vault material. When sexually dimorphic skeletal elements are not recovered, determining the sex of a decedent can be challenging. With unidentified human skeletal remains, genetic data often are evaluated in concert with anthropological analyses, as well as other types of metadata, to improve confidence in making associations or for positive identifications. This study evaluated a multi-faceted molecular genetic approach to increasing the amount of data that can be recovered from degraded skeletal remains. Results demonstrate that using a newer-generation multiplex (GlobalFiler™) with an expanded set of highly discriminatory DNA markers - combined with co-amplification of three different sex-determining loci, one additional PCR cycle, and testing multiple cuttings from the same bone or multiple regions within a skeleton - can improve reliability and accuracy in skeletal remains identifications by providing data concordance.
Bones are a valuable source of DNA in forensic, anthropological, and archaeological investigations. There are a number of scenarios in which the only samples available for testing are highly degraded and/or skeletonized. Often it is necessary to perform more than one type of marker analysis on such samples in order to compile sufficient data for identification. Lineage markers, such as Y-STRs and mitochondrial DNA (mtDNA), represent important systems to complement autosomal DNA markers and anthropological metadata in making associations between unidentified remains and living relatives or for characterization of the remains for historical and archaeological studies. In this comparative study, Y-STR typing with both Yfiler™ and Yfiler™ Plus (Thermo Fisher Scientific, Waltham, MA, USA) was performed on a variety of human skeletal remains, including samples from the American Civil War (1861-1865), the late nineteenth century gold rush era in Deadwood, SD, USA (1874-1877), the Seven Years' War (1756-1763), a seventeenth-century archaeological site in Raspenava, Bohemia (Czech Republic), and World War II (1939-1945). The skeletal remains used for this study were recovered from a wide range of environmental conditions and were extracted using several common methods. Regardless of the DNA extraction method used and the age/condition of the remains, 22 out of 24 bone samples yielded a greater number of alleles using the Yfiler™ Plus kit compared to the Yfiler™ kit using the same quantity of input DNA. There was no discernable correlation with the degradation index values for these samples. Overall, the efficacy of the Yfiler™ Plus assay was demonstrated on degraded DNA from skeletal remains. Yfiler™ Plus increases the discriminatory power over the previous generation multiplex due to the larger set of Y-STR markers available for analysis and buffer modifications with the newer version kit. Increased haplotype resolution is provided to infer or refute putative genetic relationships.
- MeSH
- alely MeSH
- DNA fingerprinting přístrojové vybavení MeSH
- kosti a kostní tkáň chemie MeSH
- lidé MeSH
- lidský chromozom Y MeSH
- mikrosatelitní repetice * MeSH
- nekrotická degradace DNA MeSH
- oběti katastrofy MeSH
- polymerázová řetězová reakce MeSH
- tělesné pozůstatky * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
AIM: A collaborative exercise with several institutes was organized by the Forensic DNA Service (FDNAS) and the Institute of the Legal Medicine, 2nd Faculty of Medicine, Charles University in Prague, Czech Republic, with the aim to test performance of different laboratories carrying out DNA analysis of relatively old bone samples. METHODS: Eighteen laboratories participating in the collaborative exercise were asked to perform DNA typing of two samples of bone powder. Two bone samples provided by the National Museum and the Institute of Archaelogy in Prague, Czech Republic, came from archeological excavations and were estimated to be approximately 150 and 400 years old. The methods of genetic characterization including autosomal, gonosomal, and mitochondrial markers was selected solely at the discretion of the participating laboratory. RESULTS: Although the participating laboratories used different extraction and amplification strategies, concordant results were obtained from the relatively intact 150 years old bone sample. Typing was more problematic with the analysis of the 400 years old bone sample due to poorer quality. CONCLUSION: The laboratories performing identification DNA analysis of bone and teeth samples should regularly test their ability to correctly perform DNA-based identification on bone samples containing degraded DNA and potential inhibitors and demonstrate that risk of contamination is minimized.
- MeSH
- DNA fingerprinting normy MeSH
- DNA analýza MeSH
- kosti a kostní tkáň chemie MeSH
- lidé MeSH
- soudní genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- multicentrická studie MeSH
- Geografické názvy
- Česká republika MeSH
Isolated populations present a constant threat to the correctness of forensic genetic casework. In this review article we present several examples of how analyzing samples from isolated populations can bias the results of the forensic statistics and analyses. We select our examples from isolated populations from central and southeastern Europe, namely the Valachs and the European Roma. We also provide the reader with general strategies and principles to improve the laboratory practice (best practice) and reporting of samples from supposedly isolated populations. These include reporting the precise population data used for computing the forensic statistics, using the appropriate θ correction factor for calculating allele frequencies, typing ancestry informative markers in samples of unknown or uncertain ethnicity and establishing ethnic-specific forensic databases.
- MeSH
- DNA fingerprinting MeSH
- etnicita genetika MeSH
- frekvence genu MeSH
- genetické markery MeSH
- genetický drift MeSH
- jednonukleotidový polymorfismus MeSH
- lidé MeSH
- lidský chromozom Y MeSH
- mikrosatelitní repetice MeSH
- mitochondriální DNA genetika MeSH
- populační genetika * MeSH
- sociální izolace * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Geografické názvy
- Evropa MeSH
Persteril 36 is a disinfectant with a broad spectrum of antimicrobial activity. Because of its bactericidal, virucidal, fungicidal, and sporicidal effectiveness, it is used as a disinfectant against biological warfare agents in the emergency and army services. In case of an attack with potentially harmful biological agents, a person's gear or afflicted skin is sprayed with a diluted solution of Persteril 36 as a precaution. Subsequently, the remains of the biological agents are analyzed. However, the question remains concerning whether DNA can be successfully analyzed from Persteril 36-treated dead bacterial cells. Spore-forming Bacillus subtilis and Gram-negative Pseudomonas aeruginosa and Xanthomonas campestris were splattered on a camouflage suit and treated with 2 or 0.2 % Persteril 36. After the disinfectant vaporized, the bacterial DNA was extracted and quantified by real-time PCR. A sufficient amount of DNA was recovered for downstream analysis only in the case of spore-forming B. subtilis treated with a 0.2 % solution of Persteril 36. The bacterial DNA was almost completely destroyed in Gram-negative bacteria or after treatment with the more concentrated solution in B. subtilis. This phenomenon can lead to false-negative results during the identification of harmful microorganisms.
- MeSH
- Bacillus subtilis genetika izolace a purifikace MeSH
- biologické bojové látky * MeSH
- dekontaminace metody MeSH
- dezinficiencia metabolismus MeSH
- diagnostické techniky molekulární metody MeSH
- DNA bakterií analýza izolace a purifikace MeSH
- fixní kombinace léků MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- kyselina peroctová metabolismus MeSH
- kyseliny sírové metabolismus MeSH
- lidé MeSH
- peroxid vodíku metabolismus MeSH
- Pseudomonas aeruginosa genetika izolace a purifikace MeSH
- senzitivita a specificita MeSH
- teoretické modely MeSH
- Xanthomonas campestris genetika izolace a purifikace MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
During a rescue excavation in October 2011, archaeologists discovered a mass grave with 10 individuals. The skeletons should belong to victims of the battle of Reichenberg between the Austrian and Prussian armies on April 21, 1757. Several bones of the skeletons were covered with a blue colored encrustation. Initial DNA analysis failed due to strong inhibition. Chemical analysis of the bluish encrustation indicated the presence of the iron phosphate mineral vivianite (Fe3(PO4)2·(H2O)8). This technical note describes a novel procedure for the removal of this inhibitory substance.
- MeSH
- DNA fingerprinting MeSH
- DNA izolace a purifikace MeSH
- fosfáty škodlivé účinky MeSH
- lidé MeSH
- odběr biologického vzorku přístrojové vybavení metody MeSH
- pohřeb MeSH
- polymerázová řetězová reakce * MeSH
- soudní antropologie přístrojové vybavení metody MeSH
- železnaté sloučeniny škodlivé účinky MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Cílem této práce je informovat o způsobu zpracování neobvyklého forenzního biologického materiálu předloženého k případu znásilnění. V rámci případu byl analyzován preparát připravený nátěrem vzorku poševního výtěru a obarvený směsí hematoxylin-eozin. Po odstranění krycího sklíčka se pomocí kapiláry mikromanipulátoru nepodařilo zajistit spermatické buňky a tyto se navíc vlivem mechanického namáhání začaly rozpadávat. Za hlavní příčinu selhání při disekci buněk považujeme pokročilou lyzi buněk. S ohledem na negativní výsledky autoři doporučují nápravná a preventivní opatření, jež zvýší šance na pozitivní identifikaci v obdobných sexuálně orientovaných případech.
The aim of this study is to provide an information about the method of processing of unusual forensic sample that was submitted for the sexual assault case. We analyzed microscopic sample of vaginal swab stained using the hematoxylin-eosin mixture. After removing the covering glass we failed to collect the sperm cells to the micromanipulator capillary. The cells even started, due to the mechanical stress, to fall apart. We think that the main reason of the microdissection failure is the advanced cell lysis. Due to the negative results of the DNA analysis we defined a set of preventive and corrective actions that would (in case of application) increase the chance of positive identification in similar sexual assault cases.
- Klíčová slova
- sexuálně orientovaný trestný čin, odběr vzorků, DNA identifikace,
- MeSH
- diagnostické techniky molekulární * metody normy přístrojové vybavení MeSH
- DNA fingerprinting * metody MeSH
- DNA analýza MeSH
- kriminologie MeSH
- lidé MeSH
- mikrodisekce MeSH
- odběr biologického vzorku * metody normy přístrojové vybavení MeSH
- soudní genetika MeSH
- spermie MeSH
- vagina chemie MeSH
- znásilnění MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- kazuistiky MeSH
