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Deoxyribonucleic Acid (DNA) hybridization is based on complementary strands of single-stranded DNA (ssDNA) hybridizing or binding to each other to form double-stranded DNA (dsDNA).

 

  • Line Probe Assay (LPA) is based on reverse hybridization between amplicons derived from a multiplex PCR and nitrocellulose-bound probes covering wild-type sequences and specific mutations in Mycobacterium tuberculosis
  • Biotin-labelled amplicons (amplified DNA of the genes of interest generated during amplification of the target DNA) are in a fluidic state. 
  • Wild-type and/ or mutated probes (reaction zones) are unlabeled and immobilized as bands onto nitrocellulose membrane strips. 
  • LPA results are based on banding patterns detected on a strip following hybridization with PCR products amplified from target DNA in clinical specimens.

 

The figure below shows the steps involved in hybridization.

 

Figure: Steps involved in Hybridization

 

 

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