Quantitation of fungal DNA contamination in commercial zymolyase and lyticase used in the preparation of fungi.
Small amounts of contaminants may lead to false-positive results in sensitive polymerase chain reaction (PCR) detection systems. To analyze contaminants and understand the usability of beta-glucanases in fungal preparations, we estimated the ribosomal DNA (rDNA) contamination in Zymolyase-100T and Lyticase by quantitative PCR. The amount of rDNA contamination determined by real-time PCR was 9210 copies/unit for Zymolyase-100T and 0.0323 copies/unit for Lyticase. The observations regarding these enzyme products indicate that careful consideration of contaminating DNA included in the reagents used for molecular diagnostics is necessary.
Miyajima
Yoshiharu
Y
Teikyo University Institute of Medical Mycology.
Satoh
Kazuo
K
Umeda
Yoshiko
Y
Makimura
Koichi
K
D004271:DNA, Fungal / Q000032:analysis*; D004275:DNA, Ribosomal / Q000032:analysis*; D004340:Drug Contamination; D005189:False Positive Reactions; D005658:Fungi / Q000235:genetics*; D004693:Glucan Endo-1,3-beta-D-Glucosidase / Q000737:chemistry*; D006026:Glycoside Hydrolases; D006867:Hydrolases / Q000737:chemistry*; D007202:Indicators and Reagents / Q000737:chemistry*; D009097:Multienzyme Complexes / Q000737:chemistry*; D010447:Peptide Hydrolases / Q000737:chemistry*; D016133:Polymerase Chain Reaction
D004271:DNA, Fungal; D004275:DNA, Ribosomal; D007202:Indicators and Reagents; D009097:Multienzyme Complexes; C025310:lyticase; C037833:zymolyase; D006867:Hydrolases; D006026:Glycoside Hydrolases; D004693:Glucan Endo-1,3-beta-D-Glucosidase; C028020:licheninase; D010447:Peptide Hydrolases