Xu Yan, Xie Jianhui, Cao Yu, Zhou Huaigu, Ping Yuan, Chen Liankang, Gu Lihua, Hu Wei, Bi Gang, Ge Jianye, Chen Xin, Zhao Ziqin
Department of Forensic Medicine, Shanghai Medical College, Fudan University, Shanghai, China; Shanghai Key Laboratory of Crime Science Evidence, Key Laboratory of Forensic Evidence and Science Technology, Ministry of Public Security, Institute of Forensic Science, Shanghai Public Security Bureau, Shanghai, China.
Department of Forensic Medicine, Shanghai Medical College, Fudan University, Shanghai, China.
PLoS One. 2014 Jul 3;9(7):e100123. doi: 10.1371/journal.pone.0100123. eCollection 2014.
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.
通过基于mRNA的分析来鉴定人体体液或组织对法医调查非常有用。先前的研究表明，mRNA生物标志物对于鉴定生物样本的来源是有效的。在本研究中，我们选择了16种组织特异性生物标志物来评估它们对人体体液和组织鉴定的特异性和敏感性，包括用于循环血液的胆色素原脱氨酶（PBGD）、血红蛋白β（HBB）和血型糖蛋白A（GLY），用于精液的鱼精蛋白2（PRM2）和转谷氨酰胺酶4（TGM4），用于阴道分泌物的粘蛋白4（MUC4）和人β-防御素1（HBD1），用于月经血的基质金属蛋白酶7和11（MMP7和MMP11），用于口腔黏膜的角蛋白4（KRT4），用于皮肤的兜甲蛋白（LOR）和胱抑素6（CST6），用于唾液的富组蛋白3（HTN3），用于鼻分泌物的富脯蛋白（STATH），用于汗液的皮肤杀菌肽（DCD）和用于尿液的尿调节素（UMOD）。上述十种常见的法医体液或组织用于评估。基于该评估，开发了一种逆转录（RT）PCR多重检测方法XCYR1，它包括12种生物标志物（即HBB、GLY、HTN3、PRM2、KRT4、MMP11、MUC4、DCD、UMOD、MMP7、TGM4和STATH）和2个管家基因[即甘油醛-3-磷酸脱氢酶（GAPDH）和18SrRNA]。该检测方法进一步用实际案件样本和模拟样本（包括单一组分和混合样本）进行了验证，并且批准XCYR1可有效鉴定法医案件样本中的常见体液或组织（即循环血液、唾液、精液、阴道分泌物、月经血、口腔黏膜、鼻分泌物、汗液和尿液）。
