SPECIFIC DETECTION OF MONKEYPOX VIRUS BY POLYMERASE-CHAIN-REACTION

The open reading frame coding for the A-type inclusion body protein (A TI) of monkeypox virus (MPV) was identified and sequenced for two stra ins. Nucleotide sequence comparison revealed 72-95.3% homology with th e reported open reading frame sequences of the ATIs of other orthopoxv irus species, such as variola, vaccinia, cowpox, ectromelia, and camel pox viruses. Each MPV strain contained an 8-bp deletion, which caused a frameshift that introduced a premature stop in the open reading fram e at base 2091 relative to the ATI open reading frame of cowpox virus strain Brighten. The sequences enabled a primer pair to be designed th at flanked the deletion and specifically amplified a 601-bp fragment t hat identified and differentiated 19 MPV strains examined from five ot her Old World orthopoxvirus species examined. The specificity was conf irmed by cleavage of the 19 MPV strain amplicons with Bg/II, which pro duced three subfragments of expected sized, based on the determined MP V sequences. (C) 1998 Elsevier Science B.V. All rights reserved.