Phylogenetic Analysis of Human Genotypes in Fars Province, Southern Iran.

BACKGROUND

This study is the first phylogenetic genotype analysis of in Iran. The main objective was to determine genotyping and identify the sub-assemblages of isolates involved in the transmission of giardiasis in Fars Province, south of Iran, in 2012.

METHODS

Forty isolates were collected from the patient's fecal samples with gastrointestinal discomfort referred to the health centers and hospitals in Shiraz, Fars Province, south of Iran. Purification of cysts from fecal samples and DNA extraction were performed using monolayer of sucrose density gradient and Phenol-Chloroform-Isoamylalcohol (PCI) respectively. Semi-nested PCR and sequence analysis were then performed using the primers (GDHeF, GDHiF, and GDHiR) which amplified a 432-bp fragment of glutamate dehydrogenase () gene. Phylogenetic analysis was carried out using a neighbor-joining tree composed of the nucleotide sequences of isolates obtained in this study and the known sequences isolates published in GenBank.

RESULTS

sub-assemblage AII was the most prevalent genotype with 80% of the cases and 20% of the cases belong to sub-assemblage BIII and BIV based on the DNA sequence of the . isolates at Fars Province were widely distributed within assemblage A cluster (sub-assemblage AII) and the remaining isolates were dispersed throughout the assemblage B cluster (sub-assemblage BIII and BIV).

CONCLUSION

PCR Sequencing and phylogenetic analysis was a proper molecular method for genotyping and discriminating of the of sub-assemblages in fecal samples, using the glutamate dehydrogenase gene that suggests a human contamination origin of giardiasis.