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. 2017:24:52.
doi: 10.1051/parasite/2017053. Epub 2017 Dec 8.

Molecular identification of Trichinella species by multiplex PCR: new insight for Trichinella murrelli

Grégory Karadjian  1 , Aurélie Heckmann  1 , Giuseppe La Rosa  2 , Edoardo Pozio  2 , Pascal Boireau  1 , Isabelle Vallée  1
Affiliations

Molecular identification of Trichinella species by multiplex PCR: new insight for Trichinella murrelli

Grégory Karadjian et al. Parasite. 2017.

Abstract
in English, French

In order to identify Trichinella at the species level, the commonly used test is a multiplex PCR, allowing the discrimination of nine out of the twelve taxa described so far. This test is based on five primer pairs amplifying fragments of the large subunit rDNA. Each taxon produces one or two bands of different sizes, resulting in a specific band pattern. By multiplex PCR, Trichinella murrelli shows two bands of 127 bp and 316 bp. However, a third band of 256 bp can occur. This band can lead to misidentification, since it is similar to the 253 bp band displayed by Trichinella britovi. BLAST analysis confirmed that the 256 bp band is from T. murrelli. The aim of this short note is to inform analysts that T. murrelli larvae may display either two- or three-band patterns.

Title: Identification moléculaire des espèces de Trichinella par PCR multiplex : nouvel éclairage pour Trichinella murrelli.

Abstract: Afin d'identifier les Trichinella au niveau de l'espèce, le test couramment utilisé est une PCR multiplex, permettant la discrimination de neuf des douze taxons décrits jusqu'à présent. Ce test est basé sur cinq paires d'amorces amplifiant des fragments de la grande sous-unité l'ADN ribosomal. Chaque taxon produit une ou deux bandes de tailles différentes, résultant en un patron de bandes spécifique. Par PCR multiplex, Trichinella murrelli présente deux bandes de 127 pb et 316 pb. Cependant, une troisième bande de 256 pb peut s'observer. Cette bande peut être la cause d'une erreur d'identification, car elle est similaire à la bande de 253 pb affichée par Trichinella britovi. L'analyse BLAST a confirmé que la bande à 256 pb provient de T. murrelli. Le but de cette note est d'informer les analystes que les larves de T. murrelli peuvent présenter des patrons à deux ou trois bandes.

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Figures

Figure 1
Figure 1
Electrophoretic profiles of Trichinella murrelli and T. britovi larva amplicons after multiplex PCR amplification.DNA extracts from 1 and 10 larvae of T. murrelli (isolate code ISS35) in lane 1 and lanes 2–4, respectively; and of T. britovi (isolate code ISS235) larva in lane 5. Lane L1 = 100 bp ladder.
Figure 2
Figure 2
Electrophoretic profiles of Trichinella murrelli uniplex PCR amplifications.DNA from T. murrelli (isolate code ISS35) reference larvae was used. Lane L = 50 bp ladder. The genes targeted were the Expansion Segment V (ESV, lane 1), Internal Transcribed Spacer 1 II (ITS1 II, Lane 2), ITS1 III (lane 3), ITS2 IV (lane 4), and ITS2 V (lane 5).
Figure 3
Figure 3
Alignment of the 256 bp fragment of ITS1 II of Trichinella murrelli obtained by uniplex PCR.BLAST analysis revealed 99.6% identity with different clones of T. murrelli, including clone 5 (Accession number KC006421).
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References

    1. Ancelle T, Dupouy-Camet J, Bougnoux ME, Fourestie V, Petit H, Mougeot G, Nozais JP, Lapierre J. 1988. Two outbreaks of trichinosis caused by horse meat in France in 1985. American Journal of Epidemiology, 127 (6), 1302­1311. - PubMed
    1. Codex Alimentarius. 2015. Guidelines for the control of Trichinella spp. in meat of suidae. CAC/GL 86–2015.
    1. Commission implementing regulation (EU) 2015/1375. 2015. Laying down specific rules on official controls for Trichinella meat. Official Journal of the European Union.
    1. Dubey JP, Hill DE, Zarlenga DS, A. 2006. Trichinella murrelli infection in a domestic dog in the United States. Veterinary Parasitology, 137(3­4), 374­378. - PubMed
    1. Dupouy-Camet J, Robert F, Guillou JP, Vallet C, Perret C, Soulé C. 1994. Identification of Trichinella isolates with random amplified polymorphic DNA markers. Parasitology Research, 80(4), 358­360. - PubMed

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