Abstract
This chapter focuses on the detection of viral pathogens using polymerase chain reaction (PCR) amplification. PCR is a relatively simple procedure consisting of repetitive cycles of oligonucleotide primer-directed DNA replication. A selected genetic target is copied in a three-step reaction: denaturing the target DNA, annealing, and extending the bound primers by DNA polymerase action. The use of Taq DNA polymerase resulted in increased specificity of PCR amplification reactions by permitting higher primer annealing and polymerase extension temperatures to be used, decreasing the amount of non-specific target amplification. The high analytical sensitivity of PCR-based procedures facilitates the detection of minute amounts of pathogens in biological specimens that are undetectable using more conventional technology. The use of uracil-N-gylcosylase in PCR protocols improves the sensitivity of PCR amplifications by degrading nonspecific extension products that result from low-stringency primer binding that occurs below the optimal primer annealing temperature. Direct detection of viral particles or nucleic acid is the most reliable method for diagnosis of human immunodeficiency virus (HIV) infection in newborns of HIV-infected women.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.