Polymerase Chain Reaction–based Differentiation of the Mosquito Sibling Species Anopheles Claviger S.S. And Anopheles Petragnani (Diptera: Culicidae)

Abstract

A polymerase chain reaction (PCR)-based diagnostic assay was developed that rapidly and reliably differentiates the sibling species of the Anopheles claviger complex, An. claviger s.s. and An. petragnani. The assay makes use of nucleotide differences in the internal transcribed spacer 2 ribosomal DNA sequences to generate PCR products of specific length for each of the two species. In evaluating the test, 580 of 592 field-collected An. claviger s.l. specimens were unambiguously identified as one of the two sibling species. Due to poor DNA quality, the remaining 12 specimens yielded no PCR product. Of the 592 mosquitoes, 407 larval specimens had been identified morphologically prior to species-specific DNA amplification, and in all instances PCR identification corroborated with morphologic identification. Mosquitoes identified as An. claviger s.s. came from various localities all over Europe and from Israel. Those identified as An. petragnani were collected in southern France and Spain. The species-diagnostic PCR assay would facilitate data collection on the temporal and spatial distribution of the two An. claviger sibling species because they represent possible vectors of disease in Europe, the Near and Middle East, and north Africa.

Malaria en España: aspectos entomológicos y perspectivas de futuro

“…Muestra estenogamia y, bajo condiciones favorables, pueden hallarse hembras activas durante los meses invernales. 34 . Presenta estenogamia, autogenia, hibernación larvaria y las hembras exhiben una clara tendencia zoofílica.…”

Ribosomal DNA second internal transcribed spacer sequence studies of Culicid vectors from an endemic area of Dirofilaria immitis in Spain

“…plumbeus, the divergence detected when comparing with the same species by Gordeev et al (2004) suggests that this Tajikistan mosquito material should be reviewed. A cryptic species might be involved in the Asian materials, as a very high number of nucleotide differences are already known between morphologically almost undifferentiable species belonging to the same complex (Kampen et al 2003). Worth mentioning is the relatively low genetic distance of An.…”

“…plumbeus (AJ555483.1) (Gordeev et al 2004); Anopheles claviger (AY129232.1) (Kampen et al 2003) and (AJ555157.1) (incomplete sequence including many indeterminate nucleotides) (Gordeev et al 2004); and Anopheles petragnani (AY129233) (Kampen et al 2003).…”

“…plumbeus from Salamanca with An. claviger from different European countries and Israel (Kampen et al 2003). This species and its sibling species An.…”

Integration of Anopheles beklemishevi (Diptera: Culicidae) in a PCR assay diagnostic for palaearctic Anopheles maculipennis sibling species

“…maculipennis (Proft et al 1999) and An. claviger complexes (Kampen et al 2003). All these assays are based on interspecific sequence differences in the ITS2 ribosomal mosquito DNA that, with the help of the PCR and species-specific primers, can be utilized to produce DNA fragments of species-specific length which are easily recognized and distinguished on an ethidium-bromide stained agarose gel.…”