Performance of 2 Polymerization Protocols Targeting Cloned Toxoplasma Parasites

Nihal A. Hanafy; Mohamed S. Badr; Ghada M. Nasr

doi:10.3889/oamjms.2018.400

Authors

Nihal A. Hanafy Department of Parasitology, Faculty of Medicine, Cairo University, Cairo, Egypt; Department of Microbiology, International Medical School, Management and Science University, Shah Alam
Mohamed S. Badr Department of Molecular Biology, Medical Research Centre, Faculty of Medicine, Ain Shams University, Cairo
Ghada M. Nasr Department of Molecular Diagnostics, Genetic Engineering and Biotechnology Research Institute, Sadat City University, Sadat City

DOI:

https://doi.org/10.3889/oamjms.2018.400

Keywords:

Toxoplasma gondii, qPCR, cPCR

Abstract

BACKGROUND: Toxoplasma gondii is a common parasitic infection of humans. Infection is usually mild. Serious complications can occur in pregnant and immunocompromised patients.

AIM: The present study aims to investigate the performance of 2 different PCR protocols; real-time quantitative molecular assays (qPCR) and conventional molecular assays (cPCR), using 2 different sets of primers and by using cloned purified Toxoplasma genomic substances to be evaluated as reference samples.

METHODS: The target DNA was provided in 8 different quantities.

RESULTS: Amplification failure was reported only with the cPCR in samples of low concentrations using both primer sets. Quantitative PCR detected the 8 different dilutions of the purified Toxoplasma gondii using the 2 sets of primers while cPCR was sensitive to detect only 6 different dilutions.

CONCLUSION: Generally real-time quantitative molecular assays, is easy to use method compared to conventional PCR assay and produces more reliable results within only one hour time but still the possible application of qPCRs in routine diagnosis necessitates analysis of a large number of clinical samples in further studies to make the proper choice.

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