Overview of Hybridization in Line Probe Assay and Genotypic Drug Sensitivity Testing

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

Resources

• GenoType MTBDR plus ver 2.0 Kit, Instructions for Use.
• GenoType MTBDRsl VER 2.0 Kit, Instructions for Use.
• GLI Training Package on LPA.

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