The Toxoplasma gondii DNA Real-Time PCR kit coded TOXODNA.CE is intended for the qualitative detection of Toxoplasma Gondii DNA in human samples (plasma and amniotic fluid) with a simultaneous control of the extraction/amplification reaction through an Internal Control (IC).
TOXODNA.CE assay was standardised against the 1st WHO International Standard for Toxoplasma gondii DNA (NIBSC code 10/242) to express positive controls concentration both in tachyzoites/ml and in International Unit (IU/ml).
The kit has been adapted for the use on the Real-Time Thermacyclers and ABI 7500 Sequence Detection System® (Software SDS version 1.3.1, Applied Biosystems™*) or MX3000P (Software MxPro version 4.01, Stratagene™***).
NTRODUCTION
The parasite Toxoplasma gondii is distributed widely in the human population, and is estimated to affect up to a 3rd of the world's population.
It is an obligate intracellular protozoan belonging to the phylum Apicomplexa, subclass Coccidia, that infects warm-blooded vertebrates, including man.
The genome is about 80Mb in size and consists of 11 chromosomes.
The most important mode of transmission of infection to humans is through the ingestion of poorly cooked meat containing encysted organism.
Farm animals in the food chain are significant reservoirs of T. gondii, which is important because of possible transmission to man, and toxoplasmosis also causes significant veterinary losses.
RINCIPLE OF THE TEST
The TOXODNA.CE Kit is based on a Real Time chemistry which uses specific Primers and Probes.
The Toxoplasma gondii DNA, recovered from the biological sample under investigation through an extraction step, is amplified using the Real Time amplification system. The amplified product is detected using a fluorescent reporter dye probe specific for a TOXO highly repeated (200 to 300 times) genomic sequence.
Heterologous Internal Control (IC) serves as an Extraction/Amplification control for each individually processed specimen aiming to the identification of reaction inhibitors.
An High Positive control (CTRL-H) and Low Positive control (CTRL-L) are supplied as controls for the PCR reaction.
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