Highlights
- •Method for the detection of a comprehensive body fluid panel using RT-LAMP with an optical detection method.
- •LAMP reaction captured as 96-well format in 30 min with a smart phone, avoiding use of expensive, complicated instrument.
- •The speed, sensitivity, and specificity this method provides is a significant improvement over current bfID methods.
Abstract
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 3 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.
Graphical abstract
Graphical Abstract
Keywords
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References
- Forensic body fluid identification: state of the art.Res. Rep. Forensic Med. Sci. 2016; 2016: 11-23
- Advanced Topics in Forensic DNA Typing.Academic Press, 2011
- The sensitivity and specificity of the RSID™-saliva kit for the detection of human salivary amylase in the Forensic Science Laboratory, Dublin, Ireland.Forensic Sci. Int. 2010; 194: 67-71
- Validation study of the Abacus diagnostics ABAcard® Hematrace® membrane test for the forensic identification of human blood.Can. Soc. Forensic Sci. J. 2003; 36: 173-183
- Prostate-specific antigen and lack of specificity for prostate cells.Lancet. 1995; 345: 1186
- Prostate specific antigen in cord blood.Indian J. Clin. Biochem. 2012; 27: 375-378
- False‐positive results with amylase testing of Citrus fruits.J. Forensic Sci. 2014; 59: 1410-1412
- Stable RNA markers for identification of blood and saliva stains revealed from whole genome expression analysis of time-wise degraded samples.Int. J. Legal Med. 2008; 122: 135-142
- Recovery and stability of RNA in vaginal swabs and blood, semen, and saliva stains.J. Forensic Sci. 2008; 53: 296-305
- Evaluation of forensic examination of extremely aged seminal stains.Leg. Med. 2014; 16: 303-307
- Forensic body fluid identification by analysis of multiple RNA markers using NanoString technology.Genomics Inform. 2013; 11: 277-281
- Evaluation of mRNA marker specificity for the identification of five human body fluids by capillary electrophoresis.Forensic Sci. Int. Genet. 2012; 6: 452-460
- Developmental validation of an enhanced mRNA-based multiplex system for body fluid and cell type identification.Sci. Justice. 2019; 59: 217-227
- mRNA profiling for body fluid identification by reverse transcription endpoint PCR and realtime PCR.Forensic Sci. Int. Genet. 2009; 3: 80-88
- rnRNA profiling for body fluid identification by reverse transcription endpoint PCR and realtime PCR.Forensic Sci. Int. Genet. 2009; 3: 80-88
- Rapid and inexpensive body fluid identification by RNA profiling-based multiplex High Resolution Melt (HRM) analysis.F1000Research. 2013; 2 (281–281)
- Facile semi-automated forensic body fluid identification by multiplex solution hybridization of NanoString® barcode probes to specific mRNA targets.Forensic Sci. Int. Genet. 2015; 14: 18-30
- A novel application of real-time RT-LAMP for body fluid identification: using HBB detection as the model.Forensic Sci. Med. Pathol. 2015; 11: 208-215
- Advances in Forensic Science: Improving Sexual Assault Evidence Analysis.University of Virginia, Charlottesville, VA2008: 184
- An extraction procedure for seminal/vaginal stains to eliminate streaking in the electrophoresis phosphoglucomutase.Crime Lab. Digest. 1985; 12: 32-33
- Rapid and sensitive salmonella typhi detection in blood and fecal samples using reverse transcription loop-mediated isothermal amplification.Foodborne Pathog. Dis. 2015; 12: 778-786
- The application of loop-mediated isothermal amplification (LAMP) in food testing for bacterial pathogens and fungal contaminants.Food Microbiol. 2013; 36: 191-206
- A valveless microfluidic device for integrated solid phase extraction and polymerase chain reaction for short tandem repeat (STR) analysis.Analyst. 2011; 136: 1928-1937
- Microarray screening and qRT-PCR evaluation of microRNA markers for forensic body fluid identification.Electrophoresis. 2014; 35: 3062-3068
- Developmental Validation of the ParaDNA(R) Screening System – a Presumptive Test for the Detection of DNA on Forensic Evidence Items.2019
- Development of HyBeacon® probes for specific mRNA detection using body fluids as a model system.Mol. Cell. Probes. 2017; 38: 51-59
- Forensic interlaboratory evaluation of the ForFLUID kit for vaginal fluids identification.J. Forensic Leg. Med. 2014; 21: 60-63
- mRNA profiling for body fluid identification by multiplex quantitative RT-PCR*.J. Forensic Sci. 2007; 52: 1252-1262
- Sensitive and rapid detection of herpes simplex virus and varicella-zoster virus DNA by loop-mediated isothermal amplification.J. Clin. Microbiol. 2005; 43: 3290-3296
- A simple identification method of saliva by detecting Streptococcus salivarius using loop‐mediated isothermal amplification.J. Forensic Sci. 2011; 56
- Shining a light on LAMP assays-A comparison of LAMP visualization methods including the novel use of berberine.Biotechniques. 2015; 58: 189-194
- Loop-mediated isothermal amplification (LAMP) of gene sequences and simple visual detection of products.Nat. Protocols. 2008; 3: 877-882
- Fast and reliable detection of Plum pox virus in woody host plants using the Blue LAMP protocol.Ann. Appl. Biol. 2011; 159: 456-466
- Evaluation of colorimetric detection methods for Shigella, Salmonella, and Vibrio cholerae by loop-mediated isothermal amplification.Diagn. Microbiol. Infect. Dis. 2013; 77: 321-323
- Colorimetric detection of loop-mediated isothermal amplification reaction by using hydroxy naphthol blue.Biotechniques. 2009; 46 (167-+)
- Visual detection of high-risk human papillomavirus genotypes 16, 18, 45, 52, and 58 by loop-mediated isothermal amplification with Hydroxynaphthol blue dye.J. Clin. Microbiol. 2011; 49: 3545-3550
- Development of Loop-Mediated Isothermal Amplification (LAMP) assay for rapid detection of Fusarium oxysporum f. sp ciceris – wilt pathogen of chickpea.BMC Res. Notes. 2015; 8
- Objective method for presumptive field-testing of illicit drug possession using centrifugal microdevices and smartphone analysis.Anal. Chem. 2016; 88: 8689-8697
- Development of a paper-based viscometer for blood plasma using colorimetric analysis.Anal. Chem. 2019; 91: 4868-4875
- Smartphone based health accessory for colorimetric detection of biomarkers in sweat and saliva.Lab Chip. 2013; 13: 3232-3238
- Smart Forensic Phone: colorimetric analysis of a bloodstain for age estimation using a smartphone.Sens. Actuators B Chem. 2017; 243: 221-225
- Validation of the loop-mediated isothermal amplification method for single nucleotide polymorphism genotyping with whole blood.Genome Lett. 2003; 2: 119-126
Article info
Publication history
Published online: November 23, 2019
Accepted:
October 25,
2019
Received in revised form:
October 4,
2019
Received:
July 24,
2019
Identification
Copyright
© 2019 Elsevier B.V. All rights reserved.
