Molecular differentiation of Hypoderma bovis and Hypoderma lineatum (Diptera, Oestridae) by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP)

The most variable region of the cytochrome oxidase I (COI) gene of Hypoderma bovis Accession number: AF497761.1 and Hypoderma lineatum Accession number: AF497762.2 (Diptera, Oestridae) was amplified by PCR and the amplicons were sequenced and analysed. PCR products were digested with three restriction enzymes, namely BfaI, HinfI and TaqI, providing informative profiles. H. bovis and H. lineatum sequences revealed an inter-specific variation rate of 8.5%, and an intra-specific variation rate of 0.87 and 0.29%, respectively. The results showed that the COI gene region examined was useful for the differentiation of H. bovis and H. lineatum and that a PCR-RFLP assay is a practical tool for their identification, offering additional diagnostic and epidemiological instruments for the study of cattle grub infestation. © 2002 Elsevier Science B.V. All rights reserved.