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