Evaluation of B1 genes nested PCR for detection Toxoplasma gondii: Comparison of six pairs of DNA primers Fitrine Ekawasti (a*), Didik T. Subekti (a), Zul Azmi (a), Muh. Ibrahim Desem (a)
a) Balai Besar Penelitian Veteriner
* fitrineekawasti[at]gmail.com
Abstract
Toxoplasmosis, caused by the obligate intracellular protozoan Toxoplasma gondii, is an important zoonosis with medical and veterinary importance worldwide. Diagnosis of toxoplasmosis has been improved by the emergence of molecular technologies to amplify parasite nucleic acids. Among these, polymerase chain reaction (PCR)-based molecular techniques have been useful for the genetic characterization of Toxoplasma gondii. PCR method has already proved to be a useful tool for the diagnosis of toxoplasmosis. This research aimed to evaluate the Primer B1 gens for PCR to diagnose T. gondii. DNA of T. gondii from liquid nitrogen was isolated using the DNAzolTM reagent. Six pairs primer of B1 genes were targeted for the development of partial length and nested PCR. Single and Nested PCR were carried out in a Thermal Cycler (Bio-Rad). PCR was evaluated for the detection of T. gondii. Each primer system used should be evaluated carefully. PCR can successfully be used to detect T. gondii and nested PCR provided higher sensitivity targeting the B1 gene for detection of Toxoplasma gondii.
