DETECTION OF HIV-1 GENOME BY TWO-STEP PCR TECHNIQUE AND OLIGONUCLEOTIDE

摘要

PURPOSE:To detect both in high specificity and sensitivity the title genome for e.g. acquired immunodeficiency syndrome (AIDS) diagnosis by determining HIV-1 (human immunodeficiency virus-1) genome using two-step PCR (polymerase chain reaction) technique. CONSTITUTION:With two-step PCR technique, using, as primer for said technique, an oligonucleotide having a gene sequence consisting of at least 15 consecutive bases among respective gene sequences of formulas I-IV with its chain length corresponding to 15-40 bases, in the first step PCR, the oligonucleotides of the formulas I and II are reacted together with a specimen and heat-resistant DNA polymerase so as to be equivalent to 25 cycles with one cycle made up of denaturation, annealing and extension. Thence, in the second step PCR, the oligonucleotides of the formulas II and III are reacted so as to be equivalent to 30 cycles in a similar way as that in the first step, thus detecting HIV-1 genome in its amplified form.