Point of care mutation detection

✍ Scribed by Mats Nilsson

• Publisher: John Wiley and Sons
• Year: 2010
• Tongue: English
• Weight: 63 KB
• Volume: 31
• Category: Article
• ISSN: 1059-7794
• DOI: 10.1002/humu.21198

✦ Synopsis

report on the development of an isothermal single-tube mutation detection technique that works directly with a drop of blood as input material. Most mutation detection techniques rely on PCR amplification, a reaction principle that for several reasons has been difficult to implement in simple distributed point-of-care diagnostics tests. One major limitation has been the construction of simple devices that permit thermal cycling as required for PCR. Also, the PCR chemistry is sensitive to components in blood, thus requiring DNA purification.

Lezhava and colleagues have developed a two-color readout of the SmartAmp2 genotyping system using so-called Exciton Primers, permitting a one-step, single-tube mutation detection reaction which delivers a result in 45 minutes directly from blood. The SmartAmp2 technique is based on an intricate exponential DNA amplification reaction utilizing hairpin structures, strand displacement polymerization, and a set of different primers. Robust genotyping is achieved by introducing a step of mismatch suppression using an enzyme that blocks synthesis of mismatched primers. The technique seems very promising for decentralized point-of-care diagnostics since the approach does not involve complicated instrumentation and sample purification.