Evaluation of sperm specific lactate dehydrogenase isoenzyme C4 (LDH C4)--application to semen detection in stains.
The usefulness of LDH C (or LDH X) as a semen-specific marker has been tested. Many different methods have been applied for identification using various electrophoretic methods such as agarose gel electrophoresis, IEF, rocket-immunoelectrophoresis with subsequent enzymatical, immunological and immunochemical detection methods. Further methods applied were dot blot analysis for LDH C and PCR analysis for X/Y-chromosome differentiation. In blood/semen mixtures the LDH C-bands were detectable in dilutions up to 50-fold after staining for total LDH activity. An appropriate extraction procedure was developed. Immunological tests using a polyclonal antibody against LDH C were found to be highly specific and highly sensitive. The immunological test gave positive results with azoospermic stains up to a few weeks and in normal sperm stains positive reactions were obtained even after 8 months storage. A comparison of different detection methods for semen showed that immunological detection methods for LDH C are, at least after longer time periods of storage, superior to microscopical detection. Swabs showed positive LDH C-reactions up to 48 h after intercourse. This reaction was closely correlated to PCR for male cells and to microscopical searching.
Pawlowski
R
R
Institut für Rechtsmedizin, Westfälische Wilhelms-Universität Münster, Federal Republic of Germany.
Brinkmann
B
B
D004587:Electrophoresis, Agar Gel / Q000592:standards; D005069:Evaluation Studies as Topic; D005554:Forensic Medicine / Q000379:methods / Q000592:standards; D006801:Humans; D015151:Immunoblotting / Q000592:standards; D007122:Immunoelectrophoresis / Q000592:standards; D007527:Isoenzymes; D007770:L-Lactate Dehydrogenase / Q000737:chemistry*; D008297:Male; D016133:Polymerase Chain Reaction / Q000592:standards; D012680:Sensitivity and Specificity; D013094:Spermatozoa / Q000737:chemistry*
D007527:Isoenzymes; D007770:L-Lactate Dehydrogenase