Polymeraise chain reaction (PCR) was the first method employed for nucleic acid amplification testing (NAAT). More recently isothermal amplification methods have been developed that utilise enzymes for DNA strand separation. An isothermal method was chosen for this project as it removes the requirement for rapid heating and cooling steps required in PCR, therefore less power is consumed within the handheld device. Isothermal amplification assays have been developed using Helicase Dependent Amplification (HDA) for Chlamydia trachomatis, Neisseria gonorrhoea, Mycoplasma genitalium and Trichmonas vaginalis with the aim to create a urethritis screening panel. The Axxin T16 was used for assay development and optimisation on the benchtop prior to performing tests on our proprietary platform.
The following shows HDA amplification curves from the proprietary isothermal amplification platform, positive and negative reactions for 1ng pCNG1.