RNA cell typing and DNA profiling of mixed samples: Can cell types and donors be associated?

Harteveld, Joyce; Lindenbergh, Alexander; Sijen, Titia

doi:10.1016/j.scijus.2013.02.001

Cited by 38 publications

(21 citation statements)

References 16 publications

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“…The mRNA-profiling multiplex technique shows promise as a method that can detect body fluids in mixed ratios [14], [46]. Since different body fluids contain different numbers of nucleated cells per unit area varying both inter- and intra- individuals, it is always useful and meaningful to choose suitable markers based on the types of body fluids mixture (differences in numbers of cells per unit volume or area).To analyze an unknown body fluid mixture, it is necessary and meaningful to first use an mRNA-profiling multiplex to find out the nature and component of the mixture.…”

Section: Discussionmentioning

confidence: 99%

Development of Highly Sensitive and Specific mRNA Multiplex System (XCYR1) for Forensic Human Body Fluids and Tissues Identification

Xie

Cao

et al. 2014

PLoS ONE

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The identification of human body fluids or tissues through mRNA-based profiling is very useful for forensic investigations. Previous studies have shown mRNA biomarkers are effective to identify the origin of biological samples. In this study, we selected 16 tissue specific biomarkers to evaluate their specificities and sensitivities for human body fluids and tissues identification, including porphobilinogen deaminase (PBGD), hemoglobin beta (HBB) and Glycophorin A (GLY) for circulatory blood, protamine 2 (PRM2) and transglutaminase 4 (TGM4) for semen, mucin 4 (MUC4) and human beta defensin 1(HBD1) for vaginal secretion, matrix metalloproteinases 7 and 11 (MMP7 and MMP11) for menstrual blood, keratin 4(KRT4) for oral mucosa, loricrin (LOR) and cystatin 6 (CST6) for skin, histatin 3(HTN3) for saliva, statherin (STATH) for nasal secretion, dermcidin (DCD) for sweat and uromodulin (UMOD) for urine. The above mentioned ten common forensic body fluids or tissues were used in the evaluation. Based on the evaluation, a reverse transcription (RT) PCR multiplex assay, XCYR1, which includes 12 biomarkers (i.e., HBB, GLY, HTN3, PRM2, KRT4, MMP11, MUC4, DCD, UMOD, MMP7, TGM4, and STATH) and 2 housekeeping genes [i.e., glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and 18SrRNA], was developed. This assay was further validated with real casework samples and mock samples (with both single source and mixture) and it was approved that XCYR1 is effective to identify common body fluids or tissues (i.e., circulatory blood, saliva, semen, vaginal secretion, menstrual blood, oral mucosa, nasal secretion, sweat and urine) in forensic casework samples.

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Section: Discussionmentioning

confidence: 99%

Development of Highly Sensitive and Specific mRNA Multiplex System (XCYR1) for Forensic Human Body Fluids and Tissues Identification

Xie

Cao

et al. 2014

PLoS ONE

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“…Samples undergoing irradiation were placed on a UVP high‐performance ultraviolet transilluminator (UVP, Upland, CA) at 302 nm for 4 h, similarly to the ultraviolet exposure of dried samples as described by Harteveld et al. . Swabs were placed directly onto the sanitized transilluminator surface and replaced into the 1.5‐mL microcentrifuge tube after treatment.…”

Section: Methodsmentioning

confidence: 99%

microRNA Detection in Blood, Urine, Semen, and Saliva Stains After Compromising Treatments

Layne

Green

Lewis

et al. 2019

Journal of Forensic Sciences

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Evaluation of microRNA (miRNA) expression as a potential method for forensic body fluid identification has been the subject of investigation over the past several years. Because of their size and encapsulation within proteins and lipids, miRNAs are inherently less susceptible to degradation than other RNAs. In this work, blood, urine, semen, and saliva were exposed to environmental and chemical conditions mimicking sample compromise at the crime scene. For many treated samples, including 100% of blood samples, miRNAs remained detectable, comparable to the untreated control. Sample degradation varied by body fluid and treatment, with blood remarkably resistant, while semen and saliva are more susceptible to environmental insult. Body fluid identification using relative miRNA expression of blood and semen of the exposed samples was 100% and 94%, respectively. Given the overall robust results herein, the case is strengthened for the use of miRNAs as a molecular method for body fluid identification.

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“…По этой же причине рекомендуется для каждого образца ставить не менее трёх воспроизведений на этапе постановки ПЦР. Специальных требований для базовой линии не существует, хотя условия ПЦР стараются подгонять таким образом, чтобы высота минимального сигнала была не ниже 150RFU, хотя некоторые опуска-ют и до 50RFU [25]. Затруднения в точности установки базовой линии, в частности, обусловлены некоторой лабильностью экспрессии генов «домашнего хозяйства» (ГДХ), амплификация которых служит своеобразным внутренним контролем [26].…”

Section: капиллярный электрофорезunclassified

“…Например, из смешанного профиля ДНК можно вычленить индивидуальные генотипы, но нередко необходимо подтверждение клеточного соответствия. В некоторых случаях, например, при типировании отпечатков, метод построения РНК-профиля может оказываться чувствительнее ДНК-типирования [10,25]. Соответственно оба метода анализа будут взаимно дополнять друг друга.…”

Section: пример анализа смешанных образцовunclassified

Tissue Typing by Means of PCR and Capillary Electrophoresis: New Aspects, Reality and Issues

Bavykin¹

2017

Russian Journal of Forensic Medicine

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ООО «Ниармедик плюс», Москва Аннотация: Целью текущего обзора является раскрытие новых перспектив для РНК-и ДНК-анализа в ру-тинной практике судебной биологии. В обзоре рассмотрены современные тканевые маркеры, основанные на матричных РНК (мРНК), методы их тестирования и подходы для совмещения РНК-анализа со стандартными протоколами исследования судебного материала. Исходя из практического опыта зарубежных исследователей, новые маркеры вполне совместимы с класси-ческими протоколами, включая использование методов ПЦР и капиллярного электрофореза, и могут быть применены после постановки иммунологических тестов, что усиливает чувствительность и не требует прин-ципиального отказа от их использования. На сегодняшний день сформированы относительно готовые мультиплексные тканевые наборы, подготовлен-ные для коммерческих реализаций в скором будущем. Популяционное генетическое разнообразие в экспрес-сии генов определяет возможность разработки отечественных панелей мРНК-маркеров на базе существующих зарубежных панелей. Ключевые слова: типирование тканей, тканевые мРНК-маркеры TISSUE TYPING BY MEANS OF PCR AND CAPILLARY ELECTROPHORESIS: NEW ASPECTS, REALITY AND ISSUESBavykin A.S. Abstract: «When the DNA is not enough» -an issue that becoming popular among forensic biologists [1]. The reasons for that are methods of tissue typing, that have limited accuracy and gradually becoming obsolete. The purpose of this review is to discuss the integration of new mRNA markers to the routine genetic forensic practice. The review discusses possibilities of utilizing the developed mRNA biomarkers for tissue typing as well as the approaches of combining gene expression analyses with standard sample workflow in forensic and criminal laboratories. Based on data obtained by the foreign researches, new mRNA markers are quite compatible with classical protocols, including PCR and capillary electrophoresis, increase the specificity of immunological methods and do not require rejection of their use. In the near future we can expect commercially developed tissue specific and multiplex assays at markets abroad. Although population diversity of genetic expression does not allow to create a universal multiplex panel, nevertheless the existing tissue mRNA panel can be enriched by the national features.

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RNA cell typing and DNA profiling of mixed samples: Can cell types and donors be associated?

Cited by 38 publications

References 16 publications

Development of Highly Sensitive and Specific mRNA Multiplex System (XCYR1) for Forensic Human Body Fluids and Tissues Identification

Development of Highly Sensitive and Specific mRNA Multiplex System (XCYR1) for Forensic Human Body Fluids and Tissues Identification

microRNA Detection in Blood, Urine, Semen, and Saliva Stains After Compromising Treatments

Tissue Typing by Means of PCR and Capillary Electrophoresis: New Aspects, Reality and Issues

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