Detection of Toxoplasma gondii by Loop-mediated Isothermal Amplification in Blood and Urine Samples from Women in Saudi Arabia

Abstract

Introduction: Toxoplasmosis is the disease which is caused by the protozoan parasite Toxoplasma gondii (T. gondii), which has the ability to infect all warm-blooded animals. Several molecular techniques for the diagnosis of Toxoplasma gondii includes normal and Real Time Polymerase Chain Reaction (RT-PCR), and gene sequencing. Aim: To apply Loop-Mediated Isothermal Amplification (LAMP) for the diagnosis of T. gondii infections in urine and blood samples from female population in Saudi Arabia. Materials and Methods: In this cross-sectional study, genomic DNA was extracted from 22 blood and urine samples (11 each), two LAMP assays based on B1 and Surface Antigen 2 (SAG2) genes of T. gondii was performed. Conventional PCR was done for the LAMP product followed by sequencing to confirm the specificity of LAMP method. Statistical Package for the Social Sciences (SPSS) version 20.0 was used to summarise continuous and categorical variables. Results: From 22 samples, 17 samples were LAMP positive in both urine and blood samples, four were negative in blood and positive in urine samples, and one was positive in blood and negative in urine. Sequencing of PCR product confirm the specificity of the method used. Conclusion: LAMP detection of T. gondii DNA is an appropriate, sensitive and specific method for diagnosis of toxoplasmosis from urine and blood specimens in humans.