Detection and subgenotyping of Cryptosporidium cuniculus in rabbit fecal samples by Real-Time PCR and high resolution melt curve analysis

• Grant number: 12/09079-6
• Support Opportunities: Scholarships abroad - Research Internship - Master's degree
• Start date: October 01, 2012
• End date: December 31, 2012
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Preventive Veterinary Medicine
• Principal Investigator: Marcelo Vasconcelos Meireles
• Grantee: Milena Sato de Souza
• Supervisor: Stephen James Hadfield
• Host Institution: Faculdade de Medicina Veterinária (FMVA). Universidade Estadual Paulista (UNESP). Campus de Araçatuba. Araçatuba , SP, Brazil
• Institution abroad: National Health Service Wales (NHS Wales), Wales
• Associated to the scholarship: 11/02730-0 - Occurrence and molecular characterization of Cryptosporidium spp. in rodents and lagomorphs raised as pets, BP.MS

Abstract

Cryptosporidium cuniculus was originally detected in rabbits and it has been identified as a human pathogen. However, the incidence, prevalence, and epidemiology of cryptosporidiosis due to C. cuniculus in rabbits and in human populations are still poorly studied. As the identification of C. cuniculus can be time consuming and expensive using the existing molecular assays, such as nested PCR followed by the sequencing of the amplified fragments, in this study it will be used real-time PCR in order to amplify fragments of the glycoprotein gp60 gene, specific for C. cuniculus Va and Vb subtypes. The real-time PCR will be performed with SYBR Green and two sets of specific primers, and after the reaction, the amplified product will be subjected to a dissociation curve analysis at high resolution (HRM). (AU)