Veronesi Fabrizia, Santoro Azzurra, Milardi Giovanni Luigi, Diaferia Manuela, Branciari Raffaella, Miraglia Dino, Cioffi Attilia, Gabrielli Simona, Ranucci David
Department of Veterinary Medicine, University of Perugia, San Costanzo Street 4, 06126, Perugia, Italy.
Department of Public Health and Infectious Diseases, Sapienza University of Rome, Aldo Moro Square 5, 00185, Rome, Italy.
Food Microbiol. 2017 May;63:213-216. doi: 10.1016/j.fm.2016.11.022. Epub 2016 Dec 2.
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.
评估了两种旨在分别靶向刚地弓形虫基因组B1区域的多拷贝序列和529 bp-RE的分子检测方法在检测猪肌肉中刚地弓形虫时的敏感性。从意大利中部集约化养殖场宰杀的498头猪身上获取膈肌柱。从组织中提取基因组DNA，并通过特定的PCR方案扩增刚地弓形虫-B1和529 bp-RE序列。在165个样本（33.13%）中检测到刚地弓形虫DNA。针对两种不同遗传标记获得的结果之间存在良好的相关性（κ = 0.77），然而，529 bp RE-PCR检测方法总体上检测到的刚地弓形虫阳性样本数量（150个样本）显著高于B1-PCR方案（134个样本）（P < 0.05）。我们的结果表明：i）应用于肌肉组织的标准化B1和529 bp-RE PCR可以检测到较高比例的刚地弓形虫感染；ii）建议采用多靶点PCR方法准确诊断猪的感染，也可用于食品检测。
