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Tuesday, July 23, 2013

Helicase Dependent Amplification...DNA Amplification Without Thermocycler


Helicase Dependent Amplification (HDA) is an invitro nucleic acid amplificatio method similar to Polymerase Chain Reaction (PCR).



HDA works at constant temperature, where as PCR requires a thermal cycler for the amplification.

Nucleic acids can be amplified by various technologies and the product can then be used in research or in diagnostics. The Polymerase Chain Reaction (PCR) dominates the nucleic acid amplification market. However, the requirement for thermocycling in PCR limits its application in certain areas, such as field-testing and point-of-care diagnostics.

Biohelix Corporation has developed Kits based on Helicase Dependent Amplification Technology.

Helicase Dependent Amplification is  an isothermal DNA amplification system that uses a helicase enzyme to unwind double stranded DNA (dsDNA), referred to as Helicase-Dependent Amplification. In HDA reactions, duplex DNA is separated into single strands by a helicase. Compared with the other techniques, this amplification method is closer to nature's method of performing DNA replication and it has the following advantages:


1. Low cost for instrumentation because use of helicase to separate DNA eliminates need for thermocycler.

2. Easy-to-use for assay development (using two primers).

3. Versatile platform: can amplify both DNA and RNA (with a reverse transcriptase) and is compatible with multiple detection technologies.

4. High sensitivity and specificity.

5. HDA is a rapid method of amplifying nucleic acid at isothermal temperature that doesn't require thermal cycler.

Disadvantages

Mass diagniosis from large no. of samples cannot be achieved.

Cost of HDA reagents are costly compared to PCR reagents.

Progress and Developments of HDA Technology

A team of researchers have successfully implemented the HDA technology for amplyfing the target DNA in combination with an integrated disposable device for DNA extraction. Full Publication can be found in the below link.

An integrated disposable device for DNA extraction and helicase dependent amplification.

http://www.ncbi.nlm.nih.gov/pubmed/20066496