Application of Molecular Diagnostic Techniques for Viral Testing

Cobo, Fernando

Ligase chain reaction assay is based on the ligation of two adjacent synthetic oligonucleotide primers which hybridize to one strand of the target DNA. A second pair of primers is used in a cycling reaction, using a thermostable DNA ligase. Both ligated products can then serve as templates for the next reaction cycle, leading to an exponential amplification process similar to PCR amplification. Detection of ligase chain reaction product could be carried out by means of several methods such as autoradiography and fluorescence. One of the advantages of the fluorescent detection system is that it is relatively easy to quantitate the amount of the ligase chain reaction product. Other methods for the detection of ligase chain reaction products in microtiter plates are been also used. LCR assays have been developed for the detection of viruses such as HPV, HSV and HIV.