Detection and identification of body fluid stains using antibody-nanoparticle conjugates.

Body fluids are considered one of the most important evidence types in forensic casework. The presence and location of blood, semen and saliva can provide crucial information to investigators. Current practice relies on an accurate visual examination followed by the use of presumptive tests to determine the identity of the body fluid type. Further laboratory based tests are required to unequivocally confirm the identity of a stain. Body fluid stains can be difficult to detect with the naked eye, particularly on dark backgrounds and hence vital evidence may be overlooked. Current methods are fluid-type specific, with a separate, and different, test required for each body fluid. The laborious nature of such analysis and the impossibility of being carried out at the crime scene, leads to a delay in the investigation process that could prove detrimental to the solving of the case. Hence, there is a need for sensitive, specific and direct methods which can simultaneously detect, differentiate, and locate human fluids on items of forensic evidence. Here, we describe the preparation of functionalized iron oxide nanoparticles conjugated to antibodies specific to blood and saliva components and their use in detecting small traces against non-contrasting substrates including glass, ceramic tile, paper and black fabric. The advantage of our technique is that it can simultaneously detect blood and saliva and can spatially locate and differentiate these body fluid types. Most importantly, our technology, which exploits the superparamagnetic properties of iron oxide nanoparticles, works in situ with no need to remove the body fluid stains for testing and with no washing steps and does not interfere with downstream DNA profiling. Thus, our technology represents a novel and effective alternative to existing methods.

[1]  M. Klintschar,et al.  Separation of sperm and vaginal cells with flow cytometry for DNA typing after sexual assault. , 1999, Obstetrics and gynecology.

[2]  Hiroyuki Honda,et al.  Medical application of functionalized magnetic nanoparticles. , 2005, Journal of bioscience and bioengineering.

[3]  S. Matsuzawa,et al.  Detection of human hemoglobin A (HbA) and human hemoglobin F (HbF) in biological stains by microtiter latex agglutination-inhibition test. , 1990, Forensic science international.

[4]  R. Sparkes,et al.  Validation of the AMPFlSTR SGM plus system for use in forensic casework. , 2000, Forensic science international.

[5]  A. Urquhart,et al.  DNA fingerprinting from single cells , 1997, Nature.

[6]  Jack Ballantyne,et al.  Multiplex mRNA profiling for the identification of body fluids. , 2005, Forensic science international.

[7]  C. Nord,et al.  A rapid method for separation and detection of human salivary amylase isoenzymes by isoelectric focusing in polyacrylamide gel. , 1976, Scandinavian journal of dental research.

[8]  Sjoerd Dirksen,et al.  Nucleophilic catalysis of hydrazone formation and transimination: implications for dynamic covalent chemistry. , 2006, Journal of the American Chemical Society.

[9]  R. Haugland,et al.  Alexa Dyes, a Series of New Fluorescent Dyes that Yield Exceptionally Bright, Photostable Conjugates , 1999, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[10]  Igor K Lednev,et al.  Raman spectroscopy offers great potential for the nondestructive confirmatory identification of body fluids. , 2008, Forensic science international.

[11]  W Bär,et al.  mRNA profiling for body fluid identification by reverse transcription endpoint PCR and realtime PCR. , 2009, Forensic science international. Genetics.

[12]  Mariaelena Pierobon,et al.  Laser capture microdissection technology , 2007, Expert review of molecular diagnostics.

[13]  B. Daniel,et al.  A novel fluorescence-based method in forensic science for the detection of blood in situ. , 2008, Forensic science international. Genetics.

[14]  K De Wael,et al.  In search of blood--detection of minute particles using spectroscopic methods. , 2008, Forensic science international.

[15]  L. I. Stowell,et al.  An enzyme-linked immunosorbent assay (ELISA) for prostate-specific antigen. , 1991, Forensic science international.

[16]  R. L. Olsen,et al.  Purification and some properties of myeloperoxidase and eosinophil peroxidase from human blood. , 1983, The Biochemical journal.

[17]  D. Thornton,et al.  Isolation and physical characterization of the MUC7 (MG2) mucin from saliva: evidence for self-association. , 1998, The Biochemical journal.

[18]  T. I. Quickenden,et al.  A study of common interferences with the forensic luminol test for blood. , 2001, Luminescence : the journal of biological and chemical luminescence.

[19]  N Kollias,et al.  A rapid method to detect dried saliva stains swabbed from human skin using fluorescence spectroscopy. , 2000, Forensic science international.

[20]  D. Willard Nanoparticles in Bioanalytics , 2003, Analytical and bioanalytical chemistry.

[21]  M. Bretscher A major protein which spans the human erythrocyte membrane. , 1971, Journal of molecular biology.

[22]  D. Sweet,et al.  PCR-based DNA typing of saliva stains recovered from human skin. , 1997, Journal of forensic sciences.

[23]  N. C. Martin,et al.  The sensitivity and specificity of red-starch paper for the detection of saliva. , 2006, Science & justice : journal of the Forensic Science Society.

[24]  R G Sweet,et al.  Fluorescence activated cell sorting. , 1972, The Review of scientific instruments.