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DNA sequence analysis suggests that cytb-nd1 PCR-RFLP may not be applicable to sandfly species identification throughout the Mediterranean region.
- Source :
-
Parasitology research [Parasitol Res] 2016 Mar; Vol. 115 (3), pp. 1287-95. Date of Electronic Publication: 2016 Jan 12. - Publication Year :
- 2016
-
Abstract
- Molecular methods are increasingly used for both species identification of sandflies and assessment of their population structure. In general, they are based on DNA sequence analysis of targets previously amplified by PCR. However, this approach requires access to DNA sequence facilities, and in some circumstances, it is time-consuming. Though DNA sequencing provides the most reliable information, other downstream PCR applications are explored to assist in species identification. Thus, it has been recently proposed that the amplification of a DNA region encompassing partially both the cytochrome-B (cytb) and the NADH dehydrogenase 1 (nd1) genes followed by RFLP analysis with the restriction enzyme Ase I allows the rapid identification of the most prevalent species of phlebotomine sandflies in the Mediterranean region. In order to confirm the suitability of this method, we collected, processed, and molecularly analyzed a total of 155 sandflies belonging to four species including Phlebotomus ariasi, P. papatasi, P. perniciosus, and Sergentomyia minuta from different regions in Spain. This data set was completed with DNA sequences available at the GenBank for species prevalent in the Mediterranean basin and the Middle East. Additionally, DNA sequences from 13 different phlebotomine species (P. ariasi, P. balcanicus, P. caucasicus, P. chabaudi, P. chadlii, P. longicuspis, P. neglectus, P. papatasi, P. perfiliewi, P. perniciosus, P. riouxi, P. sergenti, and S. minuta), from 19 countries, were added to the data set. Overall, our molecular data revealed that this PCR-RFLP method does not provide a unique and specific profile for each phlebotomine species tested. Intraspecific variability and similar RFLP patterns were frequently observed among the species tested. Our data suggest that this method may not be applicable throughout the Mediterranean region as previously proposed. Other molecular approaches like DNA barcoding or phylogenetic analyses would allow a more precise molecular species identification.
- Subjects :
- Animals
Base Sequence
Climate
DNA chemistry
DNA isolation & purification
Databases, Nucleic Acid
Female
Male
Mediterranean Region
Middle East
Phlebotomus classification
Phlebotomus genetics
Phylogeny
Polymerase Chain Reaction
Polymorphism, Restriction Fragment Length
Psychodidae genetics
Sequence Alignment
Sequence Analysis, DNA
Spain
Cytochromes b genetics
Electron Transport Complex I genetics
Psychodidae classification
Subjects
Details
- Language :
- English
- ISSN :
- 1432-1955
- Volume :
- 115
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Parasitology research
- Publication Type :
- Academic Journal
- Accession number :
- 26755361
- Full Text :
- https://doi.org/10.1007/s00436-015-4865-5