Development of HRM real-time PCR for assemblage characterization of Giardia lamblia.

Abstract A total of 90 stool samples were collected from dogs, referred to a dog shelter and a veterinary clinic. In addition, 395 stool samples obtained from pet dog owners and shelter keepers, as well as individuals referred to a medical laboratory as controls, were collected in Shahryar district, Tehran, Iran. Stool samples were parasitologically examined and the positive G. lamblia isolates were tested with Nested-PCR/sequencing for the tpi, gdh, and bg genes, and HRM real-time PCR. Microscopical examination revealed 20 (22.2%) and 34 (8.6%) Giardia-positive samples from dogs and humans, respectively. Regarding HRM real-time PCR, the prevalence of assemblages A and B in humans was 55.8% and 14.7%, respectively. In addition, 14.7% of samples were mix assemblages. HRM real-time PCR detected most of microscopically-positive samples in comparison to PCR/sequencing in both humans and dogs. The high prevalence of assemblages A and B in dogs signified the importance of a same source for infection between dogs and humans.