EVALUATION OF B1 GENE TO DETECT Toxoplasma gondii: COMPARISON OF THREE SETS NESTED PCR PRIMER

Fitrine Ekawasti, Zul Azmi, Didik Tulus Subekti, muhammad ibrahim desem, Arifin Budiman Nugraha, Siti Sa’diah, Umi Cahyaningsih

Abstract


This study aimed to evaluate three sets of B1 gene DNA primer for the diagnosis of Toxoplasma gondii. The DNA of Toxoplasma gondii that  stored on liquid nitrogen was isolated using DNAzol™ reagent. The first step of Polymerase Chain Reaction (PCRs) was performed using external and internal primer sets, respectively, and then nPCR. PCR products sequencing was performed by Apical Science. All sequences were analysed using CLC Sequence Viewer Version 8.0 software and compared to sequence database that deposited in ToxoDB (Toxoplasma gondii genome database) using BLAST (https://toxodb.org/toxo/app). Each B1 gene primer was evaluated by performing single PCR (forward and reverse) and nested PCR reactions. Three sets of B1 gene primer have different amplification precision. According to the results of amplicon sequencing, the primer set #2 has the best amplification precision of B1 gene.


Keywords


B1 gene, nested PCR, sequence, Toxoplasma gondii, zoonosis

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DOI: https://doi.org/10.21157/j.ked.hewan.v17i2.22251

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