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Molecular identification of two species of myiasis‐causing Cuterebra by multiplex PCR and RFLP
Author(s) -
Noël S.,
Tessier N.,
Angers B.,
Wood D. M.,
Lapointe F.J.
Publication year - 2004
Publication title -
medical and veterinary entomology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 82
eISSN - 1365-2915
pISSN - 0269-283X
DOI - 10.1111/j.0269-283x.2004.00489.x
Subject(s) - biology , restriction fragment length polymorphism , myiasis , zoology , intraspecific competition , polymerase chain reaction , cytochrome b , multiplex polymerase chain reaction , restriction enzyme , parasite hosting , mitochondrial dna , genetics , larva , gene , botany , world wide web , computer science
. The myiasis‐causing flies Cuterebra grisea (Coquillet) and Cuterebra fontinella (Clark) (Diptera: Oestridae) are normally parasites of mice, predominantly of the genus Peromyscus . The morphological similarities of these species and the existence of intermediate morphotypes bearing characters of both species make the identification of adults problematic; furthermore the identification of larvae is apparently not possible. This study presents two molecular approaches to discriminate between these species using specific band patterns: (i) species‐specific primers designed in the cytochrome oxidase II (COII) region used in multiplex polymerase chain reaction (PCR) and (ii) restriction fragment length polymorphism (RFLP) on amplified segments of cytochrome oxidase I (COI) gene. Both methods were tested on Cuterebra larvae and on adult museum specimens. The two techniques showed a clear difference between C. grisea and C. fontinella , although species‐specific primers were more successful than RFLP for degraded DNA. No intraspecific variation in RFLP and species‐specific amplifications were detected for the two species of Cuterebra . The results exhibit discrepancies between molecular and morphological identification, suggesting that some of the adults were misidentified.