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White light reflectance spectroscopy biosensing system for fast quantitative prostate specific antigen determination in forensic samples.

Authors
  • Koukouvinos, Georgios1
  • Metheniti, Aristea2
  • Karachaliou, Chrysoula-Evangelia3
  • Goustouridis, Dimitrios4
  • Livaniou, Evangelia3
  • Misiakos, Konstantinos5
  • Raptis, Ioannis6
  • Kondili, Aikaterini2
  • Miniati, Penelope2
  • Petrou, Panagiota7
  • Kakabakos, Sotirios8
  • 1 Immunoassay/Immunosensors Lab, Institute of Nuclear&Radiological Sciences&Technology, Energy&Safety, NCSR "Demokritos", GR-15341 Aghia Paraskevi, Greece; Forensic Science Division, Hellenic Police, Athens, Greece. , (Greece)
  • 2 Forensic Science Division, Hellenic Police, Athens, Greece. , (Greece)
  • 3 Immunopeptide chemistry Lab, Institute of Nuclear&Radiological Sciences&Technology, Energy&Safety, NCSR "Demokritos", GR-15341 Aghia Paraskevi, Greece. , (Greece)
  • 4 ThetaMetrisis S.A., Polydefkous 14, GR-12243 Athens, Greece; Electronics Eng. Dept., TEI of Piraeus, Athens, Greece. , (Greece)
  • 5 Optical sensors Lab, Institute of Nanoscience and Nanotechnology, NCSR "Demokritos", GR-15341 Aghia Paraskevi, Greece. , (Greece)
  • 6 ThetaMetrisis S.A., Polydefkous 14, GR-12243 Athens, Greece; Optical sensors Lab, Institute of Nanoscience and Nanotechnology, NCSR "Demokritos", GR-15341 Aghia Paraskevi, Greece. , (Greece)
  • 7 Immunoassay/Immunosensors Lab, Institute of Nuclear&Radiological Sciences&Technology, Energy&Safety, NCSR "Demokritos", GR-15341 Aghia Paraskevi, Greece. Electronic address: [email protected] , (Greece)
  • 8 Immunoassay/Immunosensors Lab, Institute of Nuclear&Radiological Sciences&Technology, Energy&Safety, NCSR "Demokritos", GR-15341 Aghia Paraskevi, Greece. , (Greece)
Type
Published Article
Journal
Talanta
Publication Date
Dec 01, 2017
Volume
175
Pages
443–450
Identifiers
DOI: 10.1016/j.talanta.2017.07.074
PMID: 28842014
Source
Medline
Keywords
License
Unknown

Abstract

A label-free biosensor based on white light reflectance spectroscopy for the determination of PSA as semen indicator in forensic samples is presented. The sensor is based on a two-step immunoassay which employs the same polyclonal anti-PSA antibody as capture and detection antibody followed by reaction with streptavidin as a signal enhancement step. The whole assay time was set to 10min; 5min reaction of immobilized antibody with the PSA calibrators or the samples, 3min reaction with the biotinylated anti-PSA antibody and 2min reaction with streptavidin. Following this protocol, a detection limit of 0.5ng/mL was achieved and the assay's linear response range extended up to 500ng/mL. Thus, taking into account the quantification limit of 1.0ng/mL and the average PSA concentration in semen (0.2-5.5mg/mL), semen quantities of a few nanoliters could be detected. The accuracy of the sensor developed was demonstrated through recovery (% recovery ranged from 89.6 to 106) and semen dilution experiments. A linear correlation was found for semen dilutions ranging from 5000 to 360,000. The lack of interference by other bodily fluids was confirmed by analysing stains of blood, urine and saliva prior to and after the addition of semen. Finally, the sensor was evaluated by analysing 51 forensic casework samples which were also analysed with a semi-quantitative membrane strip test (Seratec® PSA), through microscopic detection of spermatozoa, and male DNA identification through detection of Y chromosome. The results obtained with the sensor were in excellent agreement with those provided by an immunoradiometric assay kit (PSA-RIACT) and in complete agreement with the findings using the membrane strip assay, spermatozoa and Y chromosome detection. The excellent analytical performance and small size of the instrument make the sensor developed an attractive tool for use in forensic evidence screening for semen detection.

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