Liang Qiong, Sun Hongyu, Wu Xinyao, Ou Xueling, Gao Guoquan, Jin Yi, Tong Dayue
Department of Pathology, the Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou, China.
Forensic Biology Section, Sun Yat-Sen University, Guangzhou, China.
Ann Clin Lab Sci. 2018 Jan;48(1):55-62.
The aim of this study was to screen 3 mRNA markers (i.e., PAEP, LAPR3, and HOXA10) with diverse expression in different body fluids and to develop a method for the identification of menstrual blood using these mRNA markers.
Body fluid (i.e., venous blood, menstrual blood, semen, and saliva) samples were collected and prepared under differing environmental conditions (temperature, humidity and time), and RNA was extracted and reverse transcribed. The expression specificity of these markers was assessed using TaqMan probe qPCR.
A high mean cycle threshold value corresponds to a lower expression level. The mean cycle threshold value of the LAPR3, HOXA10, and PAEP genes are 8.37, 8.73, 4.67 in menstrual blood respectively. LAPR3 and PAEP were only expressed in menstrual blood. HOXA10 were expressed in blood, menstrual blood, and semen. No significant differences were found while the mean cycle threshold of MMP11 and PAEP were compared in the menstrual blood under common environmental conditions. There were no observed differences in the expression of the target genes in women of different ages and at different menstrual phases. The sensitivity of the expression of the 3 target genes could be examined in fluid amount range from 1 to 32 µl of body fluid. The expression of PAEP differed markedly from the expression of LAPR3 and HOXA10 in menstrual blood stains tested using mRNA-based assays (<0.001).
These markers, particularly PAEP, can likely be used for the identification of menstrual blood in certain forensic cases.
本研究的目的是筛选在不同体液中具有不同表达的3种mRNA标记物（即PAEP、LAPR3和HOXA10），并开发一种使用这些mRNA标记物鉴定月经血的方法。
收集体液（即静脉血、月经血、精液和唾液）样本，并在不同环境条件（温度、湿度和时间）下进行制备，提取RNA并进行逆转录。使用TaqMan探针qPCR评估这些标记物的表达特异性。
平均循环阈值越高，表达水平越低。LAPR3、HOXA10和PAEP基因在月经血中的平均循环阈值分别为8.37、8.73、4.67。LAPR3和PAEP仅在月经血中表达。HOXA10在血液、月经血和精液中均有表达。在常见环境条件下比较月经血中MMP11和PAEP的平均循环阈值时，未发现显著差异。在不同年龄和不同月经周期的女性中，未观察到目标基因表达的差异。在1至32微升体液量范围内可检测到3种目标基因表达的敏感性。在基于mRNA的检测中，月经血痕中PAEP的表达与LAPR3和HOXA10的表达明显不同（<0.001）。
这些标记物，尤其是PAEP，可能可用于某些法医案件中月经血的鉴定。
