RESUMO
Messenger RNA profiling for body fluid identification (bfID) is a useful approach to collect contextual information associated with a crime. Current methods require costly fluorescent probes, lengthy amplification protocols and/or time-consuming sample preparation. To simplify this process, we developed a bfID method that has the potential to be rapid in analysis time, inexpensive and fluorescence-free, combining a universal operating procedure with a high-throughout (microwell plate) platform for simultaneous detection of mRNA markers from whole blood, semen, saliva, and vaginal fluid. Full bfID sample preparation and analysis of 23 samples was completed in under 3â¯h using smart phone optical detection and analysis and show efficacy of the method in a validated blind study. The results provide an efficient, sensitive and specific approach to supplement the current biochemical tests in a forensic laboratory.
Assuntos
Sangue/metabolismo , Muco do Colo Uterino/metabolismo , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , RNA Mensageiro/metabolismo , Saliva/metabolismo , Sêmen/metabolismo , Smartphone , Biomarcadores/metabolismo , Feminino , Genética Forense/métodos , Globinas/genética , Globinas/metabolismo , Histatinas/genética , Histatinas/metabolismo , Humanos , Processamento de Imagem Assistida por Computador , Masculino , Proteínas Secretadas pela Vesícula Seminal/genética , Proteínas Secretadas pela Vesícula Seminal/metabolismo , Sensibilidade e Especificidade , beta-Defensinas/genética , beta-Defensinas/metabolismoRESUMO
We developed a microfluidic scanner-based profile exploration system, µSCAPE, capable of generating high resolution 3D profiles of microstructure architecture in a variety of transparent substrates. The profile is obtained by scanning the dye-filled microstructure followed by absorbance calculation and the reconstruction of the optical length at each point. The power of the method was demonstrated in (1) the inspection of the variation of the cross-section of laser-ablated PDMS channel; (2) the volume of PeT chamber; and (3) the population distribution of the volumes of the micro wells in HF-etched glass and laser-ablated PDMS. The reported methods features low equipment-cost, convenient operation and large field of view (FOV), and has revealed unreported quality parameters of the tested microstructures.
