Molecular
identification of natural sand fly species populations inferred from
ITS2 rRNA gene in Saudi Arabia
Reem A. Al-Ajmi1,
Tahany H. Ayaad1,2*, Mashael Al-Anazi3, Mohammed N.
Al-Ahdal3 and Ahmed A. Al-Qahtani3, 4
1Zoology Department,
Faculty of Science, King Saud University, Riyadh, 2457,11451, Saudi Arabia
2Entomology
Department,Faculty of Science ,Cairo University,Giza,11613,Egypt
3Department of
Infection and Immunity, Research Center, King Faisal Specialist Hospital and
Research Center, Riyadh,3354, 11211, Saudi Arabia
4Liver Disease
Research Center, King Saud University, Riyadh, 2925,11461, Saudi Arabia
*Corresponding
Author?s E-mail: ayaadmtah@gmail.com
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Publication
Data
Paper received:
10 February 2014
Revised received:
19 September 2014
Re-revised received:
29 November 2014
Accepted:
08 December 2014
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Abstract
Sandflies
are blood-sucking insects of great medical importance and are transmission vectors
for several organisms that are infectious to humans. Three hundred fourteen
adult sandflies were collected and morphologically identified as five
species: Phlebotomus papatasi, P. bergeroti, P. sergenti, Sergentomyia
antennata and S. clydei. P. papatasi was the most abundant
species (22.29%) of central region of Saudi Arabia, while S. clydei
was the most abundant species (18.79 %) of southwestern region. Molecular
classification of sandflies was done based on ribosomal RNA (rRNA) gene
Internal Transcribed Spacer 2 (ITS2) sequences. ITS2 region was amplified
using specific primers designed from a conserved region. Several clones from
each individual were sequenced and analyzed using different DNA sequence
analysis software. The results showed moderate variations (0.06%) between
clones of same P. papatasi individual. However, P. sergenti
individuals were highly divergent (0.207%). A phylogenetic tree, based on
neighbor-joining analysis, showed that morphologically identical individuals
of same species exhibited significant variations at the molecular level.
Cloned P. bergeroti sequences were highly related to P. papatasi
sequences (0.05%). Further, S. antennata species sequences were
similar to S. clydei species sequences. In conclusion, molecular typing
of sand fly species from Saudi Arabia based on ITS2 region may aid in
differentiating between species and, possibly, sub-species.
Key
words
ITS2,
Ribosomal RNA gene, Sandfly
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