Identification of screwworm species by polymerase chain reaction‐restriction fragment length polymorphism

Abstract

. 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 (F.) . Five restriction fragment length patterns were polymorphic in C. hominivorax while all fragment patterns were fixed in C . m a c e l l a r i a . 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.

Identification of Muscidifurax Spp. by Polymerase Chain Reaction–Restriction Fragment Length Polymorphism

“…However, when identifying quarantine pests, forensic samples, or in other situations in which the number of samples is limited and error, especially false negative, is unacceptable, we believe that the PCR-RFLP technique is superior. PCR-RFLP can be used to identlfy samples preserved with most commonly used preservatives as well as dried-pinned specimens (Taylor et al, 1996).…”

Population Genetics and Gene Variation of Stable Fly Populations (Diptera: Muscidae) in Nebraska

“…DNA extraction, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and restriction digest techniques were similar to Taylor et al (1996). Four amplicons were analyzed representing ' 42% of the mitochondrial genome (Table 3).…”

Application of polymerase chain reaction (PCR) in parasitology

“…Cochliomyia hominivorax και C. macellarla Η μέθοδος RFLP-PCR (Restriction fragment length polymorphism) χρησιμοποιήθηκε για το διαχωρισμό δια φόρων διπτέρων της οικογένειας Calliphoridae 43 . Ανα γνωρίστηκε και διαχωρίστηκε το είδος Cochliomyia hominivorax (Coquerel) από το C hominivorax macellarla.…”

“…Η μέθοδος δεν έχει ιδιαίτερες α παιτήσεις ως προς τη μορφή του DNA που θα χρησιμο ποιηθεί και έτσι αποφεύγονται πολλά προβλήματα συντή ρησης του δείγματος. Η RFLP-PCR είναι άμεση και φτηνή και μπορεί να αναλύσει δείγματα με απόλυτη εξειδίκευ ση σε 24 ώρες 43 .…”