Detection of Acanthamoeba From Soft Contact Lens Storage Solution by Real-time PCR

Abstract

Abstract: : Purpose: Injury and wearing of soft contact lens (SCL) are known as a cause of acanthamoeba keratitis. This study attempts to detect the subclinical presence of acanthamoeba from SCL storage solution by real-time PCR. Methods: Thirty five samples were recovered from SCL storage solution. Six samples were recovered from running water. Positive control DNA was acanthamoeba DNA which was obtained from a patient with acanthamoeba keratitis. LightCycler quick system 330 was used for amplification of target sequence. If the melting temperature of PCR products is identical to that of positive control, electrophoresis and DNA sequencing were performed. Conventional PCR was carried out to compare between the sensitivity of real-time PCR and conventional PCR. Results: Seven PCR products of 35 samples (20%) showed melting temperature identical to positive control. The sequence of the PCR product was identical to that of positive control. From running water, three of six (50%) samples were positive. The sensitivity of real-time PCR was similar to conventional PCR. Conclusion: The PCR method using LightCycler may be useful for the detection of acanthamoeba. These results indicated the presence of acanthamoeba in the SCL storage solution and running water. It is necessary to use optimal storage solution and to do proper disinfection.

Keywords: 302 Acanthamoeba • 367 contact lens • 359 clinical research methodology