title: Multilocus typing and population structure of Cryptosporidium from children in Zaragoza, Spain.
abstract: A multilocus typing approach with eight variable-number tandem-repeat (VNTR) loci and the GP60 gene was used to analyze the inter- and intra-species variation of 44 Cryptosporidium isolates from pediatric patients in Zaragoza city (NE, Spain). Restriction and sequence analyses of the SSU rRNA gene revealed that Cryptosporidium transmission is mostly anthroponotic in this area, with the predominance of Cryptosporidium hominis (n: 41) over Cryptosporidium parvum (n: 3). GP60 subtyping showed limited genetic diversity and four subtypes were identified, including IbA10G2 (n: 35), IaA24R3 (n: 6), IIaA15G1R1 (n: 1) and IIaA15G2R1 (n: 2). Five out of eight VNTR loci showed a discriminatory power higher than the GP60 gene, although each locus had a predominant allele exhibited by more than 50% of isolates. All but four alleles were associated to either C. hominis or C. parvum and linked alleles at different loci were found. Multilocus typing substantially increased the discriminatory power (Hunter-Gaston index: 0.807, 95% CI, 0.683-0.926) and revealed that genetic diversity is much higher than that reported by GP60 sequencing, since 17 multilocus subtypes (MLTs) were identified. Nearly half of the specimens were allocated to a single major MLT. However, no more than three specimens were allocated to each of the remaining MLTs. Both phylogenetic and population analyses revealed a population clustering of C. hominis according to the GP60 subtype, which indicates the robustness of this marker to differentiate genetic subpopulations. Subpopulations had an overall clonal genetic structure, although traces of genetic flow between them were also observed.
authors: Department of Animal Pathology, Faculty of Veterinary Sciences, University of Zaragoza, 50013 Zaragoza, Spain.
authors: Ana
authors: A
authors: Ramo
authors: Department of Animal Pathology, Faculty of Veterinary Sciences, University of Zaragoza, 50013 Zaragoza, Spain. Electronic address: jquilez@unizar.es.
authors: Joaquín
authors: J
authors: Quílez
authors: Department of Animal Pathology, Faculty of Veterinary Sciences, University of Zaragoza, 50013 Zaragoza, Spain.
authors: Claudia
authors: C
authors: Vergara-Castiblanco
authors: Department of Anatomy, Embryology and Genetics, Faculty of Veterinary Sciences, University of Zaragoza, 50013 Zaragoza, Spain.
authors: Luis
authors: L
authors: Monteagudo
authors: Department of Animal Pathology, Faculty of Veterinary Sciences, University of Zaragoza, 50013 Zaragoza, Spain.
authors: Emilio
authors: E
authors: Del Cacho
authors: Department of Microbiology, Preventive Medicine and Public Health, Faculty of Medicine, University of Zaragoza, 50009 Zaragoza, Spain.
authors: Antonio
authors: A
authors: Clavel
mesh_terms: D000483:Alleles; D002648:Child; D016000:Cluster Analysis; D003457:Cryptosporidiosis / Q000453:epidemiology* / Q000469:parasitology*; D003458:Cryptosporidium / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D016054:DNA, Protozoan; D005828:Genetics, Population; D006801:Humans; D018598:Minisatellite Repeats; D008969:Molecular Sequence Data; D058885:Multilocus Sequence Typing; D013030:Spain / Q000453:epidemiology
chemical_list: D016054:DNA, Protozoan
keywords: Children; Cryptosporidium; GP60 subtypes; Microsatellites; Minisatellites