An Investigation of Giardia duodenalis Subgenotype AI in Human and Cattle Isolates in Baghdad
Keywords:Giardia duodenalis, restriction fragment length polymorphism (RFLP), Glutamate dehydrogenase gene (gdh), Genotype
The aims of this study was designed for detection and determined the genotype of Giardia duodenalis isolates from human and cattle. To achieve this goal, thirty stool samples (5g) from patient were collected in sterile plastic cups, another 17 fecal samples from cattle were collected, all samples were examinedÂ microscopically by Saline wet-mount method for Giardia detection, Giardia duodenalis cysts were purified by using zinc flotation method, genomic DNA was successfully extracted from all samples by using AccuPrepÂ® Stool DNA extraction kit, the gdh gene (432 bp fragment) were amplified by using specific primers of this gene, and the restriction enzyme 2U of NIaVI was used for amplified gdh gene and found the sequence of this region for 9 human and 17 cattle sample. The results indicated that the restriction pattern showed 9samples in human isolates refer to genotype A (The digested subgenotype AI yields a fragment of 150, 120, 90bp), and 21 samples refer to genotype B (Subgenotype B allele yields an intact 432 bp fragment), while all the 17 cattle samples refer to genotype A, than result of sequencing of gdh gene and alignment showed that the 99% similarity of sample with wild type of the gdh gene of Giardia intestinalis from the Gene Bank, there are one transition at position +92 T/C single nucleotide polymorphism that cause a phenylalnine (F) to phenylalnine substitution. Conclusion: The subgenotype AI of G.duodenalisÂ was found in human and cattle isolate.
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