Practical aspects of genetic identification of hallucinogenic and other poisonous mushrooms for clinical and forensic purposes.
OBJECTIVE
To assess the usefulness of a DNA-based method for identifying mushroom species for application in forensic laboratory practice.


METHODS
Two hundred twenty-one samples of clinical forensic material (dried mushrooms, food remains, stomach contents, feces, etc) were analyzed. ITS2 region of nuclear ribosomal DNA (nrDNA) was sequenced and the sequen-ces were compared with reference sequences collected from the National Center for Biotechnology Information gene bank (GenBank). Sporological identification of mushrooms was also performed for 57 samples of clinical material.


RESULTS
Of 221 samples, positive sequencing results were obtained for 152 (69%). The highest percentage of positive results was obtained for samples of dried mushrooms (96%) and food remains (91%). Comparison with GenBank sequences enabled identification of all samples at least at the genus level. Most samples (90%) were identified at the level of species or a group of closely related species. Sporological and molecular identification were consistent at the level of species or genus for 30% of analyzed samples.


CONCLUSIONS
Molecular analysis identified a larger number of species than sporological method. It proved to be suitable for analysis of evidential material (dried hallucinogenic mushrooms) in forensic genetic laboratories as well as to complement classical methods in the analysis of clinical material.
Marek Kowalczyk, Institute of Forensic Research, Westerplatte St. 9, 31-033 Krakow, Poland, mkowalczyk@ies.krakow.pl.
Marek
M
Kowalczyk
Andrzej
A
Sekuła
Piotr
P
Mleczko
Zofia
Z
Olszowy
Anna
A
Kujawa
Szymon
S
Zubek
Tomasz
T
Kupiec
D000363:Agaricales / Q000235:genetics*; D017931:DNA Primers; D004271:DNA, Fungal / Q000032:analysis*; D004275:DNA, Ribosomal / Q000235:genetics*; D016208:Databases, Factual; D053593:Forensic Toxicology / Q000379:methods*; D006212:Hallucinations / Q000175:diagnosis*; D006213:Hallucinogens / Q000032:analysis*; D006801:Humans; D009145:Mushroom Poisoning / Q000175:diagnosis*; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D004271:DNA, Fungal; D004275:DNA, Ribosomal; D006213:Hallucinogens