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Identification of levamisole resistance markers in the parasitic nematode Haemonchus contortus using a cDNA-AFLP approach

Published online by Cambridge University Press:  03 July 2007

C. NEVEU ,
C. CHARVET ,
A. FAUVIN ,
J. CORTET ,
P. CASTAGNONE-SERENO  and
J. CABARET
C. NEVEU*
Affiliation:
INRA, IASP, 213, UR 1282, F-37380 Nouzilly, France.
C. CHARVET
Affiliation:
INRA, IASP, 213, UR 1282, F-37380 Nouzilly, France.
A. FAUVIN
Affiliation:
INRA, IASP, 213, UR 1282, F-37380 Nouzilly, France.
J. CORTET
Affiliation:
INRA, IASP, 213, UR 1282, F-37380 Nouzilly, France.
P. CASTAGNONE-SERENO
Affiliation:
INRA, UMR1064 Interactions Plantes-Microorganismes et Santé Végétale, 400 route des Chappes, BP167, 06903 Sophia Antipolis, France.
J. CABARET
Affiliation:
INRA, IASP, 213, UR 1282, F-37380 Nouzilly, France.
*
*Corresponding author: INRA, IASP, 213, UR 1282, F-37380 Nouzilly, France. Tel: +33 (0)2 47427768. Fax: +33 (0) 2 47427774. E-mail: [email protected]
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Summary

A cDNA-AFLP (cDNA-Amplified Fragment Length Polymorphism)-based strategy has been used to identify levamisole (LEV) resistance markers in the nematode Haemonchus contortus. Transcript profiles of adult nematodes from two LEV-resistant and two susceptible isolates were compared. Among the 17 280 transcript-derived fragments (TDFs) amplified, 26 presented a polymorphic pattern between resistant and susceptible nematodes: 11 TDFs were present in both resistant isolates and absent from both susceptible isolates whereas 15 TDFs were present in both susceptible isolates and absent from both resistant isolates. 8 TDFs specifically present in resistant isolates were cloned and sequenced. Some of these TDFs could represent novel genes, as their sequences presented no homologies in databases. Interestingly, specific expression of one candidate (HA17) in resistant nematodes from different isolates was confirmed by RT-PCR experiments. The finding that HA17 expression correlates with LEV resistance in three H. contortus isolates vs five susceptible isolates strongly suggest that we identified a new potential marker of LEV resistance. This differential approach at the transcriptome level could be of great interest for the identification of the molecular mechanism involved in this phenotype.

Keywords

NematodeHaemonchuslevamisoleresistancecDNA-AFLPmarker
Type
Research Article
Information
Parasitology , Volume 134 , Special Issue 8: MARKERS FOR ANTHELMINTIC RESISTANCE , August 2007 , pp. 1105 - 1110
Copyright
Copyright © Cambridge University Press 2007

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