Gene amplification using DNA from human spot urine samples.
The purpose of this study was to test the amplification of DNA from human urinary sediment for molecular epidemiological studies. Twenty-six urine samples were obtained from healthy volunteers. Polymerase chain reactions (PCR) for methylenetetrahydrofolate reductase (MTHFR), beta-globin, and N-acetyltransferase 2 (NAT2) was conducted using genomic DNA isolated from the urine. The MTHFR and beta-globin genes were amplified successfully from all the urine DNA samples while the NAT2 gene was amplified in 88.5% of cases. The median yield of DNA was 0.28 microg from the 10 ml urine samples, sufficient amounts of DNA being contained in urinary sediments for amplification of all three genes. This result indicates that urine can be used as a DNA source for PCR-based molecular epidemiological studies.
Kim
Cheong-Sik
CS
Department of Preventive Medicine, Seoul National University College of Medicine, Seoul 110-799, Republic of Korea.
Kim
Jin Hee
JH
Kang
Daehee
D
Hong
Yun-Chul
YC
Yoo
Keun-Young
KY
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001191:Arylamine N-Acetyltransferase / Q000235:genetics*; D002648:Child; D004247:DNA / Q000652:urine*; D005240:Feasibility Studies; D005260:Female; D005914:Globins / Q000235:genetics*; D006801:Humans; D008297:Male; D042965:Methylenetetrahydrofolate Reductase (NADPH2) / Q000235:genetics*; D008875:Middle Aged; D017720:Molecular Epidemiology / Q000379:methods*; D016133:Polymerase Chain Reaction; D012016:Reference Values
D005914:Globins; D004247:DNA; D042965:Methylenetetrahydrofolate Reductase (NADPH2); D001191:Arylamine N-Acetyltransferase; C478900:NAT2 protein, human