Molecular Techniques For The Detection Of Antimalarial Drug Resistance

Mutation analysis for antimalarial drug resistance currently includes a variety of strategies including PCR amplification followed by direct sequencing, single-strand conformation polymorphism (SSCP) assays, PCR followed by restriction fragment length polymorphism (RFLP) assays, allele-specific amplification assays, and real-time assays to detect point mutations conferring resistance.

In PCR-RFLP analysis, restriction enzymes are used to cut the PCR product at a specific location, proximal to the mutation site, such that wild-type and mutation-containing fragments display differing digestion patterns on agarose or polyacrylamide gel electrophoresis. SSCP is another gel mobility strategy used to detect single-point mutations in amplified DNA. PCR is used to amplify the region of interest and the resultant DNA is separated as single-stranded molecules by electrophoresis on a nondenaturing polyacrylamide gel. Wild-type and mutant single-stranded DNA fold differently, affecting their motility rate through the gel. The mutations are detected as new bands on autoradiograms (radioactive detection), by silver staining of bands or the use of fluorescent PCR primers that are subsequently detected by an automated DNA sequencer (nonradioactive detection).

Real-time PCR using hybridization probes to identify mutants based on specific melting curve patterns of the amplicon is also being developed.[30] The wealth of genetic information afforded by the completion of the Plasmodium genome will certainly help identify new drug targets, further our understanding of the molecular basis of drug resistance, and facilitate the creation of rapid assays for the detection and surveillance of drug-resistant malaria.

Getting Started With Dumbbells

Getting Started With Dumbbells

The use of dumbbells gives you a much more comprehensive strengthening effect because the workout engages your stabilizer muscles, in addition to the muscle you may be pin-pointing. Without all of the belts and artificial stabilizers of a machine, you also engage your core muscles, which are your body's natural stabilizers.

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