title: Comparison of PCR assays targeting the multi-copy targets B1 gene and 529 bp repetitive element for detection of Toxoplasma gondii in swine muscle.
abstract: The comparison of the sensitivities of two molecular assays designed to target the multi-copy sequences of the Toxoplasma gondii genomic B1 region and 529 bp-RE respectively, in detecting T. gondii in swine muscle was assessed. Diaphragm pillars were obtained from 498 slaughtered pigs managed in intensive farms in Central Italy. Genomic DNA was extracted from the tissues and T. gondii-B1 and 529 bp-RE sequences were amplified by specific PCR protocols. Toxoplasma gondii DNA was detected in 165 samples (33.13%). There was a good correlation (κ = 0.77) between the results obtained targeting the two different genetic markers, however the 529 bp RE-PCR assay overall detected a significantly higher (P < 0.05) number of T. gondii-positive samples (150 samples) than the B1-PCR protocol (134). Our results show that: i) standardized B1 and 529 bp-RE PCRs applied to muscle tissues can detect a high rate of T. gondii-infection; ii) a multi-target PCR approach is recommended for the accurate diagnosis of infection in swine and can also be used in food testing.
authors.affiliation: Department of Veterinary Medicine, University of Perugia, San Costanzo Street 4, 06126, Perugia, Italy.
authors.forename: Fabrizia
authors.initials: F
authors.lastname: Veronesi
authors.affiliation: Department of Veterinary Medicine, University of Perugia, San Costanzo Street 4, 06126, Perugia, Italy.
authors.forename: Azzurra
authors.initials: A
authors.lastname: Santoro
authors.affiliation: Department of Public Health and Infectious Diseases, Sapienza University of Rome, Aldo Moro Square 5, 00185, Rome, Italy.
authors.forename: Giovanni Luigi
authors.initials: GL
authors.lastname: Milardi
authors.affiliation: Department of Veterinary Medicine, University of Perugia, San Costanzo Street 4, 06126, Perugia, Italy.
authors.forename: Manuela
authors.initials: M
authors.lastname: Diaferia
authors.affiliation: Department of Veterinary Medicine, University of Perugia, San Costanzo Street 4, 06126, Perugia, Italy.
authors.forename: Raffaella
authors.initials: R
authors.lastname: Branciari
authors.affiliation: Department of Veterinary Medicine, University of Perugia, San Costanzo Street 4, 06126, Perugia, Italy.
authors.forename: Dino
authors.initials: D
authors.lastname: Miraglia
authors.affiliation: Unità Sanitaria Locale 1 Umbria, XX September Street 79, 06126, Perugia, Italy.
authors.forename: Attilia
authors.initials: A
authors.lastname: Cioffi
authors.affiliation: Department of Public Health and Infectious Diseases, Sapienza University of Rome, Aldo Moro Square 5, 00185, Rome, Italy.
authors.forename: Simona
authors.initials: S
authors.lastname: Gabrielli
authors.affiliation: Department of Veterinary Medicine, University of Perugia, San Costanzo Street 4, 06126, Perugia, Italy. Electronic address: david.ranucci@unipg.it.
authors.forename: David
authors.initials: D
authors.lastname: Ranucci
mesh_terms: D000818:Animals; D005516:Food Microbiology; D018503:Genome, Protozoan; D020071:Interspersed Repetitive Sequences; D007558:Italy; D009132:Muscles / Q000469:parasitology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012091:Repetitive Sequences, Nucleic Acid; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000469:parasitology; D014122:Toxoplasma / Q000235:genetics* / Q000302:isolation & purification*; D014124:Toxoplasmosis, Animal / Q000175:diagnosis* / Q000469:parasitology
keywords: 529 bp-RE; B1 gene; Muscle tissue; Swine; Toxoplasma gondii