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Identification of screwworm species by polymerase chain reaction‐restriction fragment length polymorphism
Author(s) -
TAYLOR DAVID B.,
SZALANSKI ALLEN L.,
PETERSON RICHARD D.
Publication year - 1996
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.1365-2915.1996.tb00083.x
Subject(s) - cochliomyia hominivorax , biology , restriction fragment length polymorphism , restriction enzyme , mitochondrial dna , polymerase chain reaction , restriction fragment , intraspecific competition , calliphoridae , restriction site , genetics , restriction digest , microbiology and biotechnology , dna , zoology , larva , gene , ecology
. Restriction fragment length polymorphisms in polymerase chain reaction amplified fragments (PCR‐RFLP) of mitochondrial DNA were used to differentiate species of New World screwworms (Diptera: Calliphoridae). Twenty‐seven restriction enzymes were screened on five regions of mtDNA. Eleven restriction fragment length patterns differentiated New World screwworm, Cochliomyia hominivorax (Coquerel), from secondary screwworm, Cochliomyia macellaria (R). Five restriction fragment length patterns were polymorphic in C. hominivorax while all fragment patterns were fixed in C. macellaria. Diagnostic restriction fragment length patterns were used for species diagnosis, whereas intraspecific variable patterns were used to characterize field samples and laboratory strains. The PCR‐RFLP technique is flexible with regard to developmental stage of the sample and method of preservation. We were able to characterize specimens of all life stages from egg to adult including larvae preserved in alcohol and pinned adults. PCR‐RFLP is rapid and inexpensive, enabling specimens to be characterized within 24 h for less than 2.50.