title: Development of a rapid HRM genotyping method for detection of dog-derived Giardia lamblia.
abstract: Giardia lamblia is a zoonotic flagellate protozoan in the intestine of human and many mammals including dogs. To assess a threat of dog-derived G. lamblia to humans, the common dog-derived G. lamblia assemblages A, C, and D were genotyped by high-resolution melting (HRM) technology. According to β-giardin gene sequence, the qPCR-HRM primers BG5 and BG7 were designed. A series of experiments on the stability, sensitivity, and accuracy of the HRM method were also tested. Results showed that the primers BG5 and BG7 could distinguish among three assemblages A, C, and D, which Tm value differences were about 1 °C to each other. The melting curves of intra-assay reproducibility were almost coincided, and those of inter-assay reproducibility were much the same shape. The lowest detection concentration was about 5 × 10(-6)-ng/μL sample. The genotyping results from 21 G. lamblia samples by the HRM method were in complete accordance with sequencing results. It is concluded that the HRM genotyping method is rapid, stable, specific, highly sensitive, and suitable for clinical detection and molecular epidemiological survey of dog-derived G. lamblia.
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People's Republic of China.
authors.forename: Liping
authors.initials: L
authors.lastname: Tan
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People's Republic of China.
authors.forename: Xingang
authors.initials: X
authors.lastname: Yu
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People's Republic of China.
authors.forename: Auwalu Yusuf
authors.initials: AY
authors.lastname: Abdullahi
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People's Republic of China.
authors.forename: Sheng
authors.initials: S
authors.lastname: Wu
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People's Republic of China.
authors.forename: Guochao
authors.initials: G
authors.lastname: Zheng
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People's Republic of China.
authors.forename: Wei
authors.initials: W
authors.lastname: Hu
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People's Republic of China.
authors.forename: Meiran
authors.initials: M
authors.lastname: Song
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People's Republic of China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People's Republic of China.
authors.forename: Biao
authors.initials: B
authors.lastname: Jiang
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People's Republic of China. gqli@scau.edu.cn.
authors.forename: Guoqing
authors.initials: G
authors.lastname: Li
mesh_terms: D000818:Animals; D004247:DNA / Q000737:chemistry / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004283:Dog Diseases / Q000175:diagnosis / Q000469:parasitology*; D004285:Dogs; D005243:Feces / Q000469:parasitology; D060005:Genotyping Techniques / Q000379:methods*; D016829:Giardia lamblia / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D005873:Giardiasis / Q000175:diagnosis / Q000469:parasitology / Q000662:veterinary*; D006801:Humans; D017720:Molecular Epidemiology; D044366:Transition Temperature
chemical_list: D017931:DNA Primers; D004247:DNA
keywords: Dog; Genotyping; Giardia lamblia; HRM; β-Giardin gene