Comparison of PCR assays targeting the multi-copy targets B1 gene and 529 bp repetitive element for detection of Toxoplasma gondii in swine muscle

Fabrizia Veronesi; Azzurra Santoro; Giovanni Luigi Milardi; Manuela Diaferia; Raffaella Branciari; Dino Miraglia; Attilia Cioffi; Simona Gabrielli; David Ranucci

doi:10.1016/j.fm.2016.11.022

Comparison of PCR assays targeting the multi-copy targets B1 gene and 529 bp repetitive element for detection of Toxoplasma gondii in swine muscle

Food Microbiol. 2017 May:63:213-216. doi: 10.1016/j.fm.2016.11.022. Epub 2016 Dec 2.

Authors

Fabrizia Veronesi¹, Azzurra Santoro¹, Giovanni Luigi Milardi², Manuela Diaferia¹, Raffaella Branciari¹, Dino Miraglia¹, Attilia Cioffi³, Simona Gabrielli², David Ranucci⁴

Affiliations

¹ Department of Veterinary Medicine, University of Perugia, San Costanzo Street 4, 06126, Perugia, Italy.
² Department of Public Health and Infectious Diseases, Sapienza University of Rome, Aldo Moro Square 5, 00185, Rome, Italy.
³ Unità Sanitaria Locale 1 Umbria, XX September Street 79, 06126, Perugia, Italy.
⁴ Department of Veterinary Medicine, University of Perugia, San Costanzo Street 4, 06126, Perugia, Italy. Electronic address: david.ranucci@unipg.it.

PMID: 28040171
DOI: 10.1016/j.fm.2016.11.022

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.

Keywords: 529 bp-RE; B1 gene; Muscle tissue; Swine; Toxoplasma gondii.

Publication types

Comparative Study

MeSH terms

Animals
Food Microbiology
Genome, Protozoan
Interspersed Repetitive Sequences*
Italy
Muscles / parasitology*
Polymerase Chain Reaction / methods*
Repetitive Sequences, Nucleic Acid
Sensitivity and Specificity
Swine
Swine Diseases / diagnosis*
Swine Diseases / parasitology
Toxoplasma / genetics*
Toxoplasma / isolation & purification*
Toxoplasmosis, Animal / diagnosis*
Toxoplasmosis, Animal / parasitology