Direct quantification of AD-36 adenovirus DNA by capillary electrophoresis with laser-induced fluorescence.
An adenovirus, AD-36, has been linked to human adiposity and a sensitive and reliable quantitative method is required to assess AD-36 viral loads. This report describes direct detection of AD-36 viral DNA, which is the first method to quantitate DNA without amplification. Total genomic DNA is hybridized with an AD-36 specific fluorescently labeled probe and analyzed by capillary electrophoresis with laser-induced fluorescence. The minimum detectable quantity is 10.3 ng/ml, corresponding to 282 copies of AD-36 with a precision of 1-6%. These results indicate that direct detection with capillary electrophoresis with laser-induced fluorescence (CE-LIF) is a reliable and sensitive method for quantifying AD-36 viral DNA.
Kolesar
J M
JM
School of Pharmacy, University of Wisconsin, Madison 53706, USA. jmkolesar@pharmacy.wisc.edu
Miller
J A
JA
Dhurandhar
N V
NV
Atkinson
R L
RL
D000256:Adenoviridae / Q000235:genetics*; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004279:DNA, Viral / Q000032:analysis* / Q000378:metabolism; D019075:Electrophoresis, Capillary / Q000379:methods*; D005456:Fluorescent Dyes; D007834:Lasers; D013050:Spectrometry, Fluorescence / Q000379:methods
D004279:DNA, Viral; D005456:Fluorescent Dyes; D004262:DNA Restriction Enzymes