基于组学技术的法医体液斑精准溯源的研究进展和挑战

doi:10.3969/j.issn.1671-2072.2025.06.007

摘要/Abstract

摘要： 在司法鉴定实践中，精准鉴定斑迹的体液组织来源对犯罪现场重建与案件侦破具有关键作用。系统综述了血液、唾液、精液、阴道分泌物和月经血等案件现场常见体液样本的溯源技术进展，重点探讨了包括转录组（mRNA、circRNA、miRNA和piRNA）、表观遗传组（DNA甲基化）、微生物组和蛋白质组等组学层面的精准溯源最新研究进展。同时，进一步提出引入人工智能算法、构建标准化体系、整合多组学数据以及开发新型标志物和现场检测技术的协同发展是未来研究的核心方向，为法医体液斑的精准溯源提供了系统性技术参考。

关键词: 法医物证学, 体液斑鉴定, 组学技术, 精准溯源

Abstract: In the practice of forensic appraisal, accurate identification of body fluid stains and their tissue origin plays a key role in crime scene reconstruction and case solving. A systematic review of the advances in traceability technologies for common biological samples at the scene of a case is presented, including blood, saliva, semen, vaginal secretion, menstrual blood, etc. It focuses on the potential of recent research advances in precise traceability at the omics level, including transcriptomics (mRNA, circRNA, miRNA, and piRNA), epigenetics (tissue-specific DNA methylation profiles), microbiomics, and proteomics. This study further proposes that the synergistic development of the following areas represents the core direction for future development: introducing artificial intelligence algorithms, constructing a standardized testing system, integrating multi-omics data, and developing novel markers and on-site detection techniques. This integrated approach provides a systematic technical reference for the precise traceability of forensic body fluid stains.

Key words: forensic genetics, identification of body fluid stains, omics technique, accurate traceability

中图分类号:

DF795.2

高妞, 刘晋玎, 严江伟. 基于组学技术的法医体液斑精准溯源的研究进展和挑战[J]. 中国司法鉴定, 2025(6): 60-70.

GAO Niu, LIU Jinding, YAN Jiangwei. Research Advances and Challenges of Omics Techniques for Accurate Traceability of Forensic Body Fluid Stains[J]. Chinese Journal of Forensic Sciences, 2025(6): 60-70.

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http://www.chsfjd.cn/CN/Y2025/V0/I6/60

参考文献

[ 1 ] AN J H，SHIN K J，YANG W I，et al. Body fluid identification in forensics[J]. BMB Reports，2012，45（10）：545-553.
[ 2 ] HARBISON S，FLEMING R. Forensic body fluid identification：State of the art[J]. Research and Reports in Forensic Medical Science，2016，6：11-23.
[ 3 ] VIRKLER K，LEDNEV I K. Raman spectroscopic signature of semen and its potential application to forensic body fluid identification[J]. Forensic Science International，2009，193（1/2/3）：56-62.
[ 4 ] VIRKLER K，LEDNEV I K. Forensic body fluid identification：The Raman spectroscopic signature of saliva[J]. The Analyst，2010，135（3）：512-517.
[ 5 ] VIRKLER K，LEDNEV I K. Raman spectroscopic signature of blood and its potential application to forensic body fluid identification[J]. Analytical and Bioanalytical Chemistry，2010，396（1）：525-534.
[ 6 ] SIKIRZHYTSKAYA A，SIKIRZHYTSKI V，LEDNEV I K. Raman spectroscopic signature of vaginal fluid and its potential application in forensic body fluid identification[J]. Forensic Science International，2012，216（1/2/3）：44-48.
[ 7 ] SIKIRZHYTSKI V，SIKIRZHYTSKAYA A，LEDNEV I K. Multidimensional Raman spectroscopic signature of sweat and its potential application to forensic body fluid identification[J]. Analytica Chimica Acta，2012，718：78-83.
[ 8 ] SIKIRZHYTSKI V，SIKIRZHYTSKAYA A，LEDNEV I K. Advanced statistical analysis of Raman spectroscopic data for the identification of body fluid traces：Semen and blood mixtures[J]. Forensic Science International，2012，222（1/2/3）：259-265.
[ 9 ] SIKIRZHYTSKAYA A，SIKIRZHYTSKI V，MCLAUG-HLIN G，et al. Forensic identification of blood in the presence of contaminations using Raman microspectroscopy coupled with advanced statistics：Effect of sand，dust，and soil[J]. Journal of Forensic Sciences，2013，58（5）：1141-1148.
[10] SOUKOS N S，CROWLEY K，BAMBERG M P，et al. A rapid method to detect dried saliva stains swabbed from human skin using fluorescence spectroscopy[J]. Forensic Science International，2000，114（3）：133-138.
[11] RAJ C K，GARLAPATI K，KARUNAKAR P，et al. Saliva as forensic evidence using fluorescent spectroscopy：A pilot study[J]. Journal of Clinical and Diagnostic Research，2018，12（9）：27-29.
[12] ACHETIB N，FALKENA K，SWAYAMBHU M，et al. Specific fluorescent signatures for body fluid identification using fluorescence spectroscopy[J]. Scientific Reports，2023，13：3195.
[13] 赵一霞，胡胜，叶健，等. mRNA分析在体液斑迹组织来源推断中的应用研究进展[J]. 刑事技术，2019，44（5）：388-394.
[14] HANSON E K，BALLANTYNE J. RNA profiling for the identification of the tissue origin of dried stains in forensic biology[J]. Forensic Science Review，2010，22（2）：145-157.
[15] 石珂，李淑瑾，付丽红，等. 法医体液溯源的分子生物学技术研究进展[J]. 中国法医学杂志，2019，34（3）：285-288.
[16] JUUSOLA J，BALLANTYNE J. Multiplex mRNA profiling for the identification of body fluids[J]. Forensic Science International，2005，152（1）：1-12.
[17] HAAS C，KLESSER B，MAAKE C，et al. mRNA profiling for body fluid identification by reverse transcription endpoint PCR and realtime PCR[J]. Forensic Science International：Genetics，2009，3（2）：80-88.
[18] HAAS C，HANSON E，BÄR W，et al. mRNA profiling for the identification of blood—Results of a collaborative EDNAP exercise[J]. Forensic Science International：Genetics，2011，5（1）：21-26.
[19] HAAS C，HANSON E，ANJOS M J，et al. RNA/DNA co-analysis from blood stains—Results of a second collaborative EDNAP exercise[J]. Forensic Science International： Genetics，2012，6（1）：70-80.
[20] HAAS C，HANSON E，ANJOS M J，et al. RNA/DNA co-analysis from human saliva and semen stains—Results of a third collaborative EDNAP exercise[J]. Forensic Science International： Genetics，2013，7（2）：230-239.
[21] HAAS C，HANSON E，ANJOS M J，et al. RNA/DNA co-analysis from human menstrual blood and vaginal secretion stains：Results of a fourth and fifth collaborative EDNAP exercise[J]. Forensic Science International：Genetics，2014，8（1）：203-212.
[22] HAAS C，HANSON E，BANEMANN R，et al. RNA/DNA co-analysis from human skin and contact traces—Results of a sixth collaborative EDNAP exercise[J]. Forensic Science International： Genetics，2015，16：139-147.
[23] VAN DEN BERGE M，CARRACEDO A，GOMES I，et al. A collaborative European exercise on mRNA-based body fluid/skin typing and interpretation of DNA and RNA results[J]. Forensic Science International：Genetics，2014，10：40-48.
[24] INGOLD S，DØRUM G，HANSON E，et al. Body fluid identification using a targeted mRNA massively parallel sequencing approach—Results of a EUROFORGEN/EDNAP collaborative exercise[J]. Forensic Science International： Genetics，2018，34：105-115.
[25] LAYNE T，JACKSON K，SCOTT A，et al. Optimization of novel loop-mediated isothermal amplification with colorimetric image analysis for forensic body fluid identification[J]. Journal of Forensic Sciences，2021，66（3）：1033-1041.
[26] LIU J D，ZHANG X Y，LIU Y，et al. A recombinase polymerase amplification （RPA） combined with strip visualization method for RNA-based presumptive tests of saliva and vaginal secretion[J]. Forensic Science International：Genetics，2023，62：102788.
[27] LIU J D，CHENG X J，LIU F，et al. Identification of coding region SNPs from specific and sensitive mRNA biomarkers for the deconvolution of the semen donor in a body fluid mixture[J]. Forensic Science International： Genetics，2021，52：102483.
[28] WANG S Y，JIANG T T，YUAN C Y，et al. An mRNA profiling assay incorporating coding region InDels for body fluid identification and the inference of the donor in mixed samples[J]. Forensic Science International：Genetics，2024，69：102979.
[29] LIU Z D，GAO Z，WANG J Q，et al. A method of identifying the blood contributor in mixture stains through detecting blood-specific mRNA polymorphism[J]. Electrophoresis，2020，41（15）：1364-1373.
[30] ZHANG X Y，LI J，LIU J D，et al. Identification of the vaginal secretion donor in mixture stains using polymorphic cSNPs on mRNA biomarkers[J]. Forensic Science International： Genetics，2022，59：102703.
[31] LIU Z D，WANG J Q，LI L S，et al. Identification of the body fluid donor in mixtures through target mRNA cSNP sequencing[J]. Forensic Science International：Genetics，2024，71：103066.
[32] JECK W R，SHARPLESS N E. Detecting and characterizing circular RNAs[J]. Nature Biotechnology，2014，32（5）：453-461.
[33] 张雅琪，邵诚臣，李成涛，等. circRNA的研究进展及其法医学意义[J]. 法医学杂志，2016，32（2）：131-133.
[34] ZHANG Y Q，LIU B N，SHAO C C，et al. Evaluation of the inclusion of circular RNAs in mRNA profiling in forensic body fluid identification[J]. International Journal of Legal Medicine，2018，132（1）：43-52.
[35] SONG F，LUO H B，XIE M K，et al. Microarray expression profile of circular RNAs in human body fluids[J]. Forensic Science International： Genetics，2017，6：e55-e56.
[36] LIU B N，SONG F，YANG Q R，et al. Characterization of tissue-specific biomarkers with the expression of circ-RNAs in forensically relevant body fluids[J]. International Journal of Legal Medicine，2019，133（5）：1321-1331.
[37] LIU B N，YANG Q R，MENG H，et al. Development of a multiplex system for the identification of forensically relevant body fluids[J]. Forensic Science International：Genetics，2020，47：102312.
[38] ZHAO H M，WANG C，YAO L，et al. Identification of aged bloodstains through mRNA profiling：Experiments results on selected markers of 30- and 50-year-old samples[J]. Forensic Science International，2017，272：e1-e6.
[39] HA M J，KIM V N. Regulation of microRNA biogenesis[J]. Nature Reviews Molecular Cell Biology，2014，15（8）：509-524.
[40] HANSON E K，LUBENOW H，BALLANTYNE J. Identification of forensically relevant body fluids using a panel of differentially expressed microRNAs[J]. Analytical Biochemistry，2009，387（2）：303-314.
[41] HANSON E K，REKAB K，BALLANTYNE J. Binary logistic regression models enable miRNA profiling to provide accurate identification of forensically relevant body fluids and tissues[J]. Forensic Science International：Genetics，2013，4（1）：e127-e128.
[42] ZUBAKOV D，BOERSMA A W M，CHOI Y，et al. microRNA markers for forensic body fluid identification obtained from microarray screening and quantitative RT-PCR confirmation[J]. International Journal of Legal Medicine，2010，124（3）：217-226.
[43] SAUER E，REINKE A K，COURTS C. Differentiation of five body fluids from forensic samples by expression analysis of four microRNAs using quantitative PCR[J]. Forensic Science International：Genetics，2016，22：89-99.
[44] DØRUM G，INGOLD S，HANSON E，et al. Predicting the origin of stains from whole miRNome massively parallel sequencing data[J]. Forensic Science International：Genetics，2019，40：131-139.
[45] LI Z L，LV M L，PENG D，et al. Feasibility of using probabilistic methods to analyse microRNA quantitative data in forensically relevant body fluids：A proof-of-principle study[J]. International Journal of Legal Medicine，2021，135（6）：2247-2261.
[46] WEI J W，HUANG K，YANG C，et al. Non-coding RNAs as regulators in epigenetics[J]. Oncology Reports，2017，37（1）：3-9.
[47] BAHN J H，ZHANG Q，LI F，et al. The landscape of microRNA，Piwi-interacting RNA，and circular RNA in human saliva[J]. Clinical Chemistry，2015，61（1）：221-230.
[48] RAYFORD K J，COOLEY A，RUMPH J T，et al. pi-RNAs as modulators of disease pathogenesis[J]. International Journal of Molecular Sciences，2021，22（5）：2373.
[49] YANG X，CHENG Y D，LU Q，et al. Detection of stably expressed piRNAs in human blood[J]. International Journal of Clinical and Experimental Medicine，2015，8（8）：13353-13358.
[50] WANG S Y，WANG Z，TAO R Y，et al. The potential use of Piwi-interacting RNA biomarkers in forensic body fluid identification：A proof-of-principle study[J]. Forensic Science International： Genetics，2019，39：129-135.
[51] WANG S Y，WANG Z，TAO R Y，et al. Expression profile analysis of piwi-interacting RNA in forensically relevant biological fluids[J]. Forensic Science International：Genetics，2019，42：171-180.
[52] 李文，胡胜，季安全，等. piRNAs在法医学体液鉴定中的能力评估[J]. 刑事技术，2023，48（6）：584-592.
[53] FRUMKIN D，WASSERSTROM A，BUDOWLE B，et al. DNA methylation-based forensic tissue identification[J]. Forensic Science International： Genetics，2011，5（5）：517-524.
[54] PARK J L，KWON O H，KIM J H，et al. Identification of body fluid-specific DNA methylation markers for use in forensic science[J]. Forensic Science International： Genetics，2014，13：147-153.
[55] LEE H Y，AN J H，JUNG S E，et al. Genome-wide methylation profiling and a multiplex construction for the identification of body fluids using epigenetic markers[J]. Forensic Science International： Genetics，2015，17：17-24.
[56] LEE H Y，JUNG S E，LEE E H，et al. DNA methylation profiling for a confirmatory test for blood，saliva，semen，vaginal fluid and menstrual blood[J]. Forensic Science International： Genetics，2016，24：75-82.
[57] LI Z Q，LIU N，YUAN F，et al. Development of a novel panel for blood identification based on blood-specific CpG-linked SNP markers[J]. International Journal of Legal Medicine，2024，138（3）：1205-1219.
[58] LI Z Q，LI J T，LI Y D，et al. Development of a multiplex methylation-sensitive restriction enzyme-based SNP typing system for deconvolution of semen-containing mixtures[J]. International Journal of Legal Medicine，2021，135（4）：1281-1294.
[59] LI Z Q，LI Y D，LIU N，et al. Typing of semen-containing mixtures using ARMS-based semen-specific CpG-InDel/STR markers[J]. International Journal of Legal Medicine，2022，136（4）：1163-1176.
[60] 中国抗癌协会肿瘤标志专业委员会. 肿瘤DNA甲基化标志物检测及临床应用专家共识（2024版）[J]. 中国癌症防治杂志，2024，16（2）：129-142.
[61] AN J H，CHOI A，SHIN K J，et al. DNA methylation-specific multiplex assays for body fluid identification[J]. International Journal of Legal Medicine，2013，127（1）：35-43.
[62] BETSER-COHEN G，MAYUONI-KIRSEHENBAUM L，ZUBEIDAT K，et al. Identification of oral bacteria as a new forensic tool for saliva detection[J]. Forensic Science International，2024，361：112112.
[63] HUANG H Y，YAO T，WU W B，et al. Specific microbes of saliva and vaginal fluid of Guangdong Han females based on 16S rDNA high-throughput sequencing[J]. International Journal of Legal Medicine，2019，133（3）：699-710.
[64] DOBAY A，HAAS C，FUCILE G，et al. Microbiome-based body fluid identification of samples exposed to indoor conditions[J]. Forensic Science International： Genetics，2019，40：105-113.
[65] NAKANISHI H，SHOJO H，OHMORI T，et al. Identification of feces by detection of Bacteroides genes[J]. Forensic Science International： Genetics，2013，7（1）：176-179.
[66] ZHENG Y Y，YE L Y，DU J Y，et al. Changes in the microbial community of semen exposed to different simulated forensic situations[J]. Microbiology Spectrum，2024，12（8）：e0012524.
[67] DÍEZ LÓPEZ C，VIDAKI A，RALF A，et al. Novel taxonomy-independent deep learning microbiome approach allows for accurate classification of different forensically relevant human epithelial materials[J]. Forensic Science International： Genetics，2019，41：72-82.
[68] WOHLFAHRT D，TAN-TORRES A L，GREEN R，et al. A bacterial signature-based method for the identification of seven forensically relevant human body fluids[J]. Forensic Science International： Genetics，2023，65：102865.
[69] YU D J，WANG T，ZHANG L W，et al. Identification of body fluid sources based on microbiome antibiotic resistance genes using high-throughput qPCR[J]. Forensic Science International： Genetics，2025，77：103241.
[70] HU S，LOO J A，WONG D T. Human body fluid proteome analysis[J]. Proteomics，2006，6（23）：6326-6353.
[71] LI S J，PENG M，LI H，et al. Sys-BodyFluid：A systematical database for human body fluid proteome research[J]. Nucleic Acids Research，2009，37：D907-D912.
[72] VAN STEENDAM K，DE CEULENEER M，DHAENENS M，et al. Mass spectrometry-based proteomics as a tool to identify biological matrices in forensic science[J]. International Journal of Legal Medicine，2013，127（2）：287-298.
[73] YANG H Y，ZHOU B，DENG H T，et al. Body fluid identification by mass spectrometry[J]. International Journal of Legal Medicine，2013，127（6）：1065-1077.
[74] LEGG K M，POWELL R，REISDORPH N，et al. Discovery of highly specific protein markers for the identification of biological stains[J]. Electrophoresis，2014，35（21/22）：3069-3078.
[75] LEGG K M，POWELL R，REISDORPH N，et al. Verification of protein biomarker specificity for the identification of biological stains by quadrupole time-of-flight mass spectrometry[J]. Electrophoresis，2017，38（6）：833-845.
[76] KAMANNA S，HENRY J，VOELCKER N H，et al. Direct identification of forensic body fluids using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry[J]. International Journal of Mass Spectrometry，2016，397/398：18-26.
[77] KUSHNER I K，CLAIR G，PURVINE S O，et al. Individual variability of protein expression in human tissues[J]. Journal of Proteome Research，2018，17（11）：3914-3922.
[78] CHOI A，SHIN K J，YANG W I，et al. Body fluid identification by integrated analysis of DNA methylation and body fluid-specific microbial DNA[J]. International Journal of Legal Medicine，2014，128（1）：33-41.
[79] ZUBAKOV D，CHAMIER-CIEMIŃSKA J，KOKMEIJER I，et al. Introducing novel type of human DNA markers for forensic tissue identification：DNA copy number variation allows the detection of blood and semen[J]. Forensic Science International： Genetics，2018，36：112-118.
[80] JIANG Y M，SUN J，HUANG X，et al. Direct identification of forensic body fluids by MALDI-MS[J]. Analyst，2019，144（23）：7017-7023.