LIMITATIONS OF ELISA IMMUNOLOGICAL ANALYSES IN DETERMINING THE POSTMORTEM INTERVAL
I. Turlea, G. Curca, L. Matei, C. Trandafir, F. Perde
Abstract: Introduction. The evaluation of postmortem interval, PMI, is of great medico-legal and judicial importance. Quantitative and qualitative immunological methods can be used, such as ELISA and Western Blot, immunohistochemistry, peptidome methods (e.g. creatine kinase, myoglobin and heat shock proteins for PMI), metabolomic methods (e.g. quantification of putrescine and cadaverine by GC-MS and LC-MS in PMI), biophysical methods, FTIR (Fourier Transform Infrared Spectroscopy highlighting postmortem changes in the structure of proteins in the vitreous humour in PMI).
Material and methods. To determine PMI, we imagined a pilot study to apply ELISA (Enzyme-Linked Immunosorbent Assay) method to search protein extract from fragments of lateral vastus quadriceps muscle. We used the ELISA “sandwich” line in the department of immunology of Serology Laboratory in the National Institute of Legal Medicine Mina Minovici Bucharest, a semi-automatic line composed of a shaking incubator, ELISA microplate washer and PR1400 Bio-Rad microplate reader with specific software (Magellan).
Results. Our aim is to present advantages of ELISA in forensic use and some limiting laboratory aspects and possible solutions (i.e. irregularities in the postmortem variations of the tested markers, the need to identify useful samples for validation).
Conclusions. ELISA as a complementary analyse of degradation of proteins of interest can be successfully used in forensic practice to determine medical causes of death and PMI. Keywords: ELISA, serology, limits, postmortem interval, time since death.
, metabolomic methods (e.g. quantification of putrescine and cadaverine by GC-MS and LC-MS in PMI), biophysical methods, FTIR (Fourier Transform Infrared Spectroscopy highlighting postmortem changes in the structure of proteins in the vitreous humour in PMI).
<br /> Material and methods. To determine PMI, we imagined a pilot study to apply ELISA (Enzyme-Linked Immunosorbent Assay) method to search protein extract from fragments of lateral vastus quadriceps muscle. We used the ELISA “sandwich” line in the department of immunology of Serology Laboratory in the National Institute of Legal Medicine Mina Minovici Bucharest, a semi-automatic line composed of a shaking incubator, ELISA microplate washer and PR1400 Bio-Rad microplate reader with specific software (Magellan).
<br /> Results. Our aim is to present advantages of ELISA in forensic use and some limiting laboratory aspects and possible solutions (i.e. irregularities in the postmortem variations of the tested markers, the need to identify useful samples for validation).
<br /> Conclusions. ELISA as a complementary analyse of degradation of proteins of interest can be successfully used in forensic practice to determine medical causes of death and PMI.&redirect_uri=http://rjlm.ro/index.php/arhiv/1052)