Real-time PCR assay for detection of Trichinella in meat

被引:10
作者
Alonso, Mercedes [1 ]
Herrero, Beatriz [1 ]
Vieites, Juan M. [1 ]
Espineira, Montserrat [1 ]
机构
[1] ANFACO CECOPESCA, Area Mol Biol & Biotechnol, Vigo 36310, Pontevedra, Spain
关键词
Trichinella; Detection; Meat; TaqMan; Real-time PCR; PCR; POLYMERASE-CHAIN-REACTION; QUANTITATIVE PCR; ENZYME-IMMUNOASSAY; MULTIPLEX PCR; PARASITIC NEMATODES; INTERGENIC REGION; IDENTIFICATION; TRICHINOSIS; INFECTION; DNA;
D O I
10.1016/j.foodcont.2011.02.009
中图分类号
TS2 [食品工业];
学科分类号
0832 ;
摘要
Trichinella are a group of widely distributed parasites which have acquired high social relevance due to their involvement in foodborne infections caused by consumption of raw or undercooked food. A TaqMan(R)-LNA probe Real-time PCR assay targeting the 5S rRNA was developed allowing the simultaneous detection of the 10 species and 3 genotypes of Trichinella present in meat tissues. The detection limit employing dilutions of genomic DNA was 2 pg and the determination of the detection limt in terms of ppm was 1 ppm. The main novelty of this work lies in the fact that it can assure the absence of 10 species and 3 genotypes of Trichinella in an only assay. The proposed methodology is rapid, robust, highly sensitive and readily adaptable in routine molecular diagnostic laboratories, and can be employed as molecular screening method in order to assess the food security. (C) 2011 Elsevier Ltd. All rights reserved.
引用
收藏
页码:1333 / 1338
页数:6
相关论文
共 54 条
[1]   RANDOM AMPLIFIED POLYMORPHIC DNA FINGERPRINTS OF THE 8 TAXA OF TRICHINELLA AND THEIR COMPARISON WITH ALLOZYME ANALYSIS [J].
BANDI, C ;
LAROSA, G ;
BARDIN, MG ;
DAMIANI, G ;
COMINCINI, S ;
TASCIOTTI, L ;
POZIO, E .
PARASITOLOGY, 1995, 110 :401-407
[2]   A real-time PCR assay for quantifying Plasmodium falciparum infections in the mosquito vector [J].
Bell, AS ;
Ranford-Cartwright, LC .
INTERNATIONAL JOURNAL FOR PARASITOLOGY, 2004, 34 (07) :795-802
[3]  
Bell AS, 2002, TRENDS PARASITOL, V18, P337, DOI 10.1016/S1471-4922(02)02331-0
[4]   Use of mitochondrial RNA genes for the differentiation of four Trichinella species by multiplex PCR amplification [J].
Blaga, R. ;
Fu, BaoQuan ;
Le Rhun, D. ;
Le Naour, E. ;
Heckman, A. ;
Zocevic, A. ;
Liu, MingYuan ;
Boireau, P. .
JOURNAL OF HELMINTHOLOGY, 2009, 83 (02) :121-128
[5]   Trichinella in horses:: a low frequency infection with high human risk [J].
Boireau, P ;
Vallée, I ;
Roman, T ;
Perret, C ;
Liu, MY ;
Gamble, HR ;
Gajadhar, A .
VETERINARY PARASITOLOGY, 2000, 93 (3-4) :309-320
[6]   Molecular identification of Trichinella spiralis and Trichinella britovi by diagnostic multiprimer large mitochondrial rRNA amplification [J].
Borsuk, P ;
Moskwa, B ;
Pastusiak, K ;
Cabaj, W .
PARASITOLOGY RESEARCH, 2003, 91 (05) :374-377
[7]  
BUSTIN SA, 2002, J MOL ENDOCRINOLOGY, V29
[8]   Simple species identification of Trichinella isolates by amplification and sequencing of the 5S ribosomal DNA intergenic spacer region [J].
De Bruyne, A ;
Yera, H ;
Le Guerhier, F ;
Boireau, P ;
Dupouy-Camet, J .
VETERINARY PARASITOLOGY, 2005, 132 (1-2) :57-61
[9]   Trichinellosis: a worldwide zoonosis [J].
Dupouy-Camet, J .
VETERINARY PARASITOLOGY, 2000, 93 (3-4) :191-200
[10]  
*EC, 2005, OFFICIAL J EURO 1222, P66