Limits...
Sandfly fever Sicilian virus, Algeria.

Izri A, Temmam S, Moureau G, Hamrioui B, de Lamballerie X, Charrel RN - Emerging Infect. Dis. (2008)

Bottom Line: A sequence closely related to that of SFSV was detected in a Phlebotomus ariasi sandfly.Of 60 human serum samples, 3 contained immunoglobulin G against SFSV.These data suggest SFSV is present in Algeria.

View Article: PubMed Central - PubMed

Affiliation: Université Paris 13, Bobigny, France.

ABSTRACT
To determine whether sandfly fever Sicilian virus (SFSV) is present in Algeria, we tested sandflies for phlebovirus RNA. A sequence closely related to that of SFSV was detected in a Phlebotomus ariasi sandfly. Of 60 human serum samples, 3 contained immunoglobulin G against SFSV. These data suggest SFSV is present in Algeria.

Show MeSH

Related in: MedlinePlus

Map of Algeria showing where sandflies were trapped (â– ).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2600228&req=5

Figure 1: Map of Algeria showing where sandflies were trapped (â– ).

Mentions: Over a 4-night period in July 2006, a total of 460 sandflies were trapped as described (12). Trapping was performed at Larbaa Nath Iraten (previously known as Fort National) in the Kabylian region of Algeria, near Tizi Ouzout (Figure 1). CDC Miniature Light Traps were adapted for sandfly capture by using an ultrafine mesh. Traps were hung 1–2 m above ground. They were placed during late afternoon in or near animal housing facilities (chickens, rabbits, goats, horses). Each morning, sandflies were collected, identified morphologically, and placed in 1.5-mL microfuge tubes. Captured sandflies belonged to 7 species: P. perniciosus (n = 364), P. longicuspis (n = 61), P. sergenti (n = 21), P. ariasi (n = 6), P. perfiliewi (n = 3), P. papatasi (n = 1), and Sergentomyia minuta (n = 1). They were organized into 24 pools, each containing up to 30 sandflies. Each pool was ground in RNA NOW chaotropic solution (Ozyme, Montigny le Bretonneux, France). RNA purification was performed according to the manufacturer’s protocol. A total of 10 μL of RNA suspension was used for reverse transcription with random hexanucleotide primers with the Taqman Reverse Transcription Reagents (Applied Biosystems, Foster City, CA, USA) in a final volume of 50 μL, according to the manufacturer’s recommended protocol. To test these specimens for Toscana virus RNA and phlebovirus RNA, we used 10 μL of cDNA in the previously described assays (12,13). Pool F tested positive with nested primers Phlebo2+/Phlebo2–. The PCR product was sequenced directly with primers used for PCR amplification. A 201-nt sequence (excluding primers) was obtained and submitted to the National Center for Biotechnology Information BLAST program, which retrieved a unique hit, consisting of Cyprus phlebovirus polymerase gene with a 92% identity score. Because Cyprus virus has been reported to be closely related to SFSV (2), we amplified and sequenced the homologous genome region of SFSV strain Sabin by using the primers described above. The same approach was applied to Arbia virus, a related phlebovirus isolated in Italy simultaneously with Toscana virus for comparative analysis. These 3 sequences were deposited in the GenBank database under accession nos. EU240880, EU266619, and EU266620. Laboratory contamination can be excluded because the sequence corresponding to SFSV-Algeria is divergent from its closest sequence (SFSV-Italy-Sabin) by 4%, which corresponds to 8 nt mutations; in addition, SFSV-Italy-Sabin has been manipulated after PCR amplification and sequencing of SFSV-Algeria to compare it genetically with the sequence obtained from Algerian sandflies.


Sandfly fever Sicilian virus, Algeria.

Izri A, Temmam S, Moureau G, Hamrioui B, de Lamballerie X, Charrel RN - Emerging Infect. Dis. (2008)

Map of Algeria showing where sandflies were trapped (â– ).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2600228&req=5

Figure 1: Map of Algeria showing where sandflies were trapped (â– ).
Mentions: Over a 4-night period in July 2006, a total of 460 sandflies were trapped as described (12). Trapping was performed at Larbaa Nath Iraten (previously known as Fort National) in the Kabylian region of Algeria, near Tizi Ouzout (Figure 1). CDC Miniature Light Traps were adapted for sandfly capture by using an ultrafine mesh. Traps were hung 1–2 m above ground. They were placed during late afternoon in or near animal housing facilities (chickens, rabbits, goats, horses). Each morning, sandflies were collected, identified morphologically, and placed in 1.5-mL microfuge tubes. Captured sandflies belonged to 7 species: P. perniciosus (n = 364), P. longicuspis (n = 61), P. sergenti (n = 21), P. ariasi (n = 6), P. perfiliewi (n = 3), P. papatasi (n = 1), and Sergentomyia minuta (n = 1). They were organized into 24 pools, each containing up to 30 sandflies. Each pool was ground in RNA NOW chaotropic solution (Ozyme, Montigny le Bretonneux, France). RNA purification was performed according to the manufacturer’s protocol. A total of 10 μL of RNA suspension was used for reverse transcription with random hexanucleotide primers with the Taqman Reverse Transcription Reagents (Applied Biosystems, Foster City, CA, USA) in a final volume of 50 μL, according to the manufacturer’s recommended protocol. To test these specimens for Toscana virus RNA and phlebovirus RNA, we used 10 μL of cDNA in the previously described assays (12,13). Pool F tested positive with nested primers Phlebo2+/Phlebo2–. The PCR product was sequenced directly with primers used for PCR amplification. A 201-nt sequence (excluding primers) was obtained and submitted to the National Center for Biotechnology Information BLAST program, which retrieved a unique hit, consisting of Cyprus phlebovirus polymerase gene with a 92% identity score. Because Cyprus virus has been reported to be closely related to SFSV (2), we amplified and sequenced the homologous genome region of SFSV strain Sabin by using the primers described above. The same approach was applied to Arbia virus, a related phlebovirus isolated in Italy simultaneously with Toscana virus for comparative analysis. These 3 sequences were deposited in the GenBank database under accession nos. EU240880, EU266619, and EU266620. Laboratory contamination can be excluded because the sequence corresponding to SFSV-Algeria is divergent from its closest sequence (SFSV-Italy-Sabin) by 4%, which corresponds to 8 nt mutations; in addition, SFSV-Italy-Sabin has been manipulated after PCR amplification and sequencing of SFSV-Algeria to compare it genetically with the sequence obtained from Algerian sandflies.

Bottom Line: A sequence closely related to that of SFSV was detected in a Phlebotomus ariasi sandfly.Of 60 human serum samples, 3 contained immunoglobulin G against SFSV.These data suggest SFSV is present in Algeria.

View Article: PubMed Central - PubMed

Affiliation: Université Paris 13, Bobigny, France.

ABSTRACT
To determine whether sandfly fever Sicilian virus (SFSV) is present in Algeria, we tested sandflies for phlebovirus RNA. A sequence closely related to that of SFSV was detected in a Phlebotomus ariasi sandfly. Of 60 human serum samples, 3 contained immunoglobulin G against SFSV. These data suggest SFSV is present in Algeria.

Show MeSH
Related in: MedlinePlus