SENSITIVE AND COST-EFFECTIVE DETECTION OF TOXOPLASMA GONDII IN WATER SUPPLIES OF THE BLACK SEA IN TURKEY BY LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (LAMP)

Abstract

Detection of Toxoplasma in water samples was performed by loop-mediated isothermal amplification (LAMP) assay of Toxoplasma gondii B I gene and nested polymerase chain reaction assays (PCR) of the 18S-rRNA gene. The identification limit (sensitivity) of T gondii LAMP assay is 100 fg/mu L of T gondii DNA. Thirty randomly selected water pellets were spiked with 10 Toxoplasma oocysts to illustrate the sensitivity of LAMP and nested PCR. When the LAMP assay results were compared with those from the nested PCR, the identification sensitivity in spiked pellets was 100% by LAMP and 43.33% by nested PCR. Furthermore, 60 natural water samples of different sources were outright analyzed by LAMP and nested PCR. Twenty-five percent (15/60) and 10% (6/60) of the natural water samples were positive for Toxoplasma DNA by LAMP and nested PCR, respectively These data indicate that Toxoplasma contamination can be detected in water samples of Black Sea area in Turkey by LAMP and nested PCR methods. Biotechnol. & Biotechnol. Eq. 2013, 27(1), 3543-3546