Abstract Molecular techniques are of growing in the study of anthelmintic resistance in trichostrongylid worm population.One of these techniques is PCR –RFLP that have recently developed for detection of BZ resistance in Haemonchus contortus and Teladorsagia circumcincta. In this study for the determination of resistance at position 200 of β-tubulin isotype I, we compare PCR –RFLP(using TaaI) with an innovative restriction site created PCR-RFLP (using PSP1). In the first step, the genomic DNA was isolated from single Teladorsagia circumcincta(30) and Haemonchus contortus(30) collected from the abomasums of untreated sheep.Only Teladosagia circumcincta collected from the abomasums of treated sheep(20) with 5 mg/Kg BW 2.5% albendazole suspension.These were amplified with the primers pair creating a 403bp PCR products in length. In the second step, the PCR products were extracted from agarose gel and amplified with the modified forward primers (UTvetMF-primers) and the same reverse primers as in step 1, creating a PCR products of 222bp. The PCR products were then cut with PSP1 to obtain in the case of normal allele two DNA products (183bp and 39bp).Also this products were digested with TaaI which cuts mutated allele into two fragment. Fifteen of Teladorsagia circumcincta collected from treated sheep and twenty of Teladorsagia circumcincta from untreated sheep cuted with PSP1406I that were BZRS heterozygote . Five of Teladorsagia circumcinta in treated sheep were not cut by PSP1406I that have been BZRR homozygote. Ten of Teladorsagia circumcinta untreated sheep were BZSS homozygote because cuted complately into two fragment with PSP1406I.None of these worms in both groups was not cuted with TaaI even after 16 h incubation in 65 °C. All of Haemonchus contortus that collected from untreated sheep was complately cuted with PSP1406I because all of Haemonchus contortus were BZSS homozygote and was not cuted by TaaI. In our preliminary experiment we could not find BZRR homozygote form of Haemonchus contortus within the examined samples.

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