Agudelo Juliana, Huynh Crystal, Halámek Jan
Department of Chemistry, University at Albany, SUNY, Albany NY 12222, USA.
Analyst. 2015 Mar 7;140(5):1411-5. doi: 10.1039/c4an02269f. Epub 2015 Jan 23.
A bioaffinity-driven cascade assay was developed to determine the time elapsed from the point a blood sample was left at a crime scene to the point of discovery. Two blood markers, creatine kinase (CK) and alanine transaminase (ALT), were utilized to determine the age of the blood spot based on their natural denaturation processes. The analysis with the proposed bioassay was performed in human serum samples, which underwent the aging process under environmental conditions that could be expected at crime scenes. The concentration of the markers in the sample was based on physiological levels present in healthy adults. These two markers were concerted in a biocatalytic cascade composed of two parallel subsystems, with each of them following the activity of one marker. Both markers have very distinct denaturation rates which would not allow them to be used in a single marker setup while still providing satisfactory results. However, by parallel tunable monitoring of both markers, it is possible to provide information of the blood sample age with low temporal error for a prolonged period of time. To mimic a possible real crime scene situation – the reliability of the proposed assay was then successfully tested on dried/aged serum samples (up to 5 days old) in environments with different temperatures.
开发了一种生物亲和力驱动的级联检测方法，以确定从血样留在犯罪现场到被发现时所经过的时间。利用两种血液标志物，即肌酸激酶（CK）和丙氨酸转氨酶（ALT），根据它们的自然变性过程来确定血斑的时间。使用所提出的生物检测方法对人血清样本进行分析，这些样本在犯罪现场可能出现的环境条件下经历老化过程。样本中标志物的浓度基于健康成年人的生理水平。这两种标志物在由两个平行子系统组成的生物催化级联中协同作用，每个子系统跟踪一种标志物的活性。两种标志物具有非常不同的变性速率，这使得它们不能在单一标志物设置中使用，同时仍能提供令人满意的结果。然而，通过对两种标志物进行并行可调监测，有可能在较长时间内以低时间误差提供血样时间信息。为了模拟可能的真实犯罪现场情况，然后在不同温度环境下对干燥/老化的血清样本（长达5天）成功测试了所提出检测方法的可靠性。
