Detection of adenovirus using PCR and molecular beacon

The polymerase chain reaction (PCR) and a molecular beacon probe were used for the detection of Adenovirus. A 307 bp DNA fragment from a conserved reg ion of the hexon gene was amplified. The specific molecular beacon was char acterized with respect to its efficiency of quenching, and signal to noise ratio by spectrofluorometric analysis of its hybridization with virus specf ic complementary single stranded oligonucleotide target. Amplification was carried out in the presence of the molecular beacon probe, and the amplifie d target was detected by measurement of fluorescence signal in the post PCR sample. Separately, a P-32-labeled linear probe (having the same sequence as that of molecular beacon probe) was liquid-phase hybridized with the pro duct of PCR performed in the absence of the molecular beacon. The virus spe cific target was then detected by electrophoresis of the hybridized product in a nondenaturing polyacrylamide gel and subsequent autoradiographic anal ysis. The detection limit of adenovirus by PCR in the presence of the molec ular beacon probe was found to be similar to that obtained by labeled linea r probe hybridization following PCR. (C) 1999 Elsevier Science B.V. All rig hts reserved.