After cycling is completed, the amplification products can be examined in various ways. Typically, the contents of the reaction vessel are subjected to gel electrophoresis. Additional product analysis by probe hybridization or direct DNA sequencing is often performed to further verify the authenticity of the amplicon. Additional probes hybridize to the viral RNA, which then hybridize to the bDNA amplifiers. Alkaline phosphatase-labeled probes then hybridize to the bDNA allowing for amplification of the signal.
Detection is accomplished using chemiluminescence technology.
Reference to an external standard curve permits quantitation. Oligonucleotide pairs hybridize to target sequences within the gene or the cryptic plasmid. The bound oligonucleotides are separated by a small gap at the target site. The enzyme DNA polymerase uses nucleotides in the LCR reaction mixture to fill in this gap, creating a ligatable junction. Once the gap is filled, DNA ligase joins the oligonucleotide pairs to form a short single-stranded product that is complementary to the original target sequence.