Summary
The possibility of typing DNA polymorphisms on urine samples was investigated in a controlled storage experiment and for samples that were 1-7 years old. Female urine samples showed a higher amount of epithelial cells and therefore a higher DNA yield. Employing the polymerase chain reaction, specific amplification results were achieved for all samples over a 6 month storage period. The microscopical examination of the samples revealed not only differing degrees of contamination with bacteria, yeasts and fungi, but also the presence of still intact epithelial cells. Only 20% of the male samples and 32% of the female samples yielded specific amplification results. By separating the human cells from the contaminating organisms prior to DNA extraction, the number of successfully typed samples could be improved to 35% of the male and 77% of the female samples. This result confirms that excess amounts of coextracted non-human DNA can inhibit the specific amplification of human target sequences.
Zusammenfassung
Die Typisierbarkeit von DNA-Polymorphismen an gelagerten Urinproben wurde in einem systematischen Lagerungsexperiment und stichprobenartig au 1-7 Jahre alten Proben untersucht. In beiden Untersuchungen zeigte sich bei Frauen eine erhöhte Zahl von Epithelzellen im Urin und damit eine größere Menge an extrahierbarer DNA. Mit dem Verfahren der Genamplifikation konnten für den untersuchten Zeitraum von sechs Monaten alle Proben typisiert werden. Die mikroskopische Begutachtung der mehrere Jahre alten Proben zeigte verschiedene Grade von Kontamination mit Bakterien, Hefen und Pilzen, aber auch das Vorhandensein von intakten Epithelzellen. Nur 20% der Urinproben von Männern und 32% der Proben von Frauen ergaben spezifische Amplifikationsergebnisse. Durch die Trennung von humanen Zellen und kontaminierenden Organismen vor der DNA-Extraktion konnte diese Quote auf 35% der männlichen und 77% der weiblichen Proben erhöht werden. Dieses Ergebnis zeigt, daß ein hoher Anteil koextrahierter nicht menschlicher DNA einen hemmenden Effekt auf die spezifische Amplifikation humaner Zielsequenzen hat.
Similar content being viewed by others
References
Allen RC, Graves G, Budowle B (1989) Polymerase chain reaction amplification products separated on rehydratable polyacrylamide gels and stained with silver. Bio Techniques 7:736–744
Boerwinkle E, Xiong W, Fourest E, Chan L (1989) Rapid typing of tandemly repeated hypervariable loci by the polymerase chain reaction: application to the apolipoprotein B 3′ hypervariable region. Proc Natl Acad Sci USA 86:212–216
Brinkmann B (1992) The use of STR's in stain analysis. Proceedings 3rd International Symposium Human Identification, Promega, Madison, pp 357–373
Brinkmann B, Rand S, Bajanowski T (1992) Forensic identification of urine samples. Int J Leg Med 105:59–61
Chase MG (1986) ABO typing studies on liquid urines. J Forensic Sci 31:881–885
Gasparini P, Savoia A, Pignatti PF, Dallapiccola B, Novelli G (1989) Amplification of DNA from epithelial cells in urine. New Engl J Med 320:809
Gibb B (1965) Vergleichende Untersuchung zur Ausscheidung gruppengeprägter Substanzen des ABO(H)-Systems im Speichel und Urin. Z Arztl Fortb 59:185–189
Gill P, Jeffreys AJ, Werrett DJ (1985) Forensic application of DNA-fingerprints. Nature 318:577–579
Gyllenstein UB, Josefsson A, Schemschat K, Saldeen T, Petterson U (1992) DNA typing of material with mixed genotypes using allele-specific enzymatic amplification (polymerase chain reaction). Forensic Sci Int 52:149–160
Mills PR, Chase MG (1989) The detection of group specific component from urine samples. Forensic Sci Int 43:215–221
Pötsch L, Prager-Eberle M, Penzes L, Schneider P, Rittner C (1993) Zum Identitätsnachweis an Urinproben mit PCR. Rechtsmed 3:128–133
Rand S, Puers C, Skowasch K, Wiegand P, Budowle B, Brinkmann B (1992) Population genetics and forensic efficiency data of 4 AMPFLP's. Int J Leg Med 104:329–333
Roewer L, Nürnberg P, Fuhrmann E, Rose M, Prokop O, Epplen JT (1990) Stain analysis using oligonucleotide probes specific for simple repetitive DNA sequences. Forensic Sci Int 47:59–70
Ruano G, Brash DE, Kidd KK (1991) PCR: The first few cycles. Amplifications 7:1–4
Singer-Sam J, Tanguay RL, Riggs AD (1989) Use of chelex to improve the PCR signal from a small number of cells. Amplifications 3:11
Stefan RJ, Atlas RM (1988) DNA amplification to enhance detection of genetically engineered bacteria in environmental samples. Appl Environ Microbiol 54:2185–2191
Waye JS, Michaud D, Bowen JH, Fourney RM (1991) Sensitive and specific quantification of human genomic deoxyribonucleic acid (DNA) in forensic science specimens: casework examples. J Forensic Sci 36:1198–1203
Author information
Authors and Affiliations
Additional information
Dedicated to Prof. Dr. M. Staak on the occasion of his 60th birthday
Rights and permissions
About this article
Cite this article
Prinz, M., Grellner, W. & Schmitt, C. DNA typing of urine samples following several years of storage. Int J Leg Med 106, 75–79 (1993). https://doi.org/10.1007/BF01225044
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01225044