Random Amplification of Polymorphic DNA as a Tool for Taxonomic Studies of Triatomine Bugs (Hemiptera: Reduviidae)

Abstract

Eleven of 27 decameric primers were found to be suitable for random amplification of polymorphic DNA (RAPD) from triatomine bugs on the basis that they produced discrete profiles and distinguished among Panstrongylus megistus (Burmeister), Rhodnius prolixus Stål, and Triatoma infestans (Klug). The legs, or single leg segments, of individual bugs were used as the source of DNA so that the taxonomic value of the bug was conserved. Within the scope of the specimens studied, RAPD profiles allowed assignment to species even when bugs were kept dry for up to 12 mo. Profiles for individuals within a species were not identical. RAPD profiles, with the specimens tested, distinguished among species of 3 pairs considered to be morphologically similar and closely related, namely, Rhodnius ecuadorensis Lent & León and Rhodnius pictipes Stål; Rhodnius nasutus Stål, and Rhodnius neglectus Lent; Rhodnius prolixus Stål and Rhodnius robustus Larrousse. RAPD data conformed with the perceived affinities among these species. RAPD polymorphisms were seen with T. infestans from 3 different localities, but none of the polymorphisms was confined to 1 source. RAPD provided a molecular basis to reassess taxonomic relationships within the Triatomine subfamily. The accurate distinction of triatomine species and of intraspecific bug populations may contribute to elimination of vector-borne Chagas disease from the Americas.

Genetic polymorphism, molecular characterization and relatedness of Macrobrachium species (Palaemonidae) based on RAPD-PCR

“…Among the methodologies available, the RAPD-PCR technique has been amply utilized in different organisms, due to its aforementioned characteristics, with results considered important for this objective (Garcia et al, 1998;Iqbal et al, 2007;Camargo Jr. et al, 2007;Roratto et al, 2008;Guerra Jr. et al, 2010).…”

“…Among the many molecular methods currently available for genetic studies, it appears particularly suitable for analysis of any species, revealing a high degree of polymorphism in many cases (Williams et al, 1990;Garcia et al, 1998;Castiglioni and de Campos Bicudo, 2005;Roratto et al, 2008;Cesniene et al, 2010;Nath et al, 2010). The method uses PCR with a single short oligonucleotide primer that randomly amplifies short fragments of genomic DNA, separated by size, with agarose gel electrophoresis.…”

Genetic Relationships Among <I>Aedes aegypti</I> (Diptera: Culicidae) Populations from Argentina Using Random Amplified Polymorphic DNA Polymerase Chain Reaction Markers

“…Molecular markers generated by RAPD-PCR were useful in determining the geographic origins of a weevil (Williams et al 1994), stored product moth (Dowdy and McGaughey 1996), gypsy moth (Schreiber et al 1997), and fruit flies (Reyes and Onchado 1998), and in estimating gene flow and genetic variability in C. hominivorax (InfanteMalachias et al 1999;Azeredo-Espin and Lessinger 2006) as well as several species of medically-important mosquitoes (de Sousa et al 2001;Gonzáles et al 2007;Hiragi et al 2009). Additionally, inter-and intraspecific differentiation by RAPD-PCR has been applied to the Mediterranean fruit fly (Sonvico et al 1996), triatomine bugs (Garcia et al 1998), and horn flies (Castiglioni et al 2005), with the capability of identifying biotypes or ecotypes/ecoraces of various insect species (Guirao et al 1997;Pornkulwat et al 1998;Saha and Kundu 2006). Here, our objective was to further develop RAPD-PCR for accurate and timely identification of C. hominivorax from other wound inhabiting flies, and to gain insight into the potential of RAPD-PCR to discriminate geographic origin of screwworm samples.…”

The number of families of Triatoma dimidiata in a Guatemalan house

“…RAPD-PCR was performed with the four RAPD primers that had previously been shown to amplify triatomine DNA (Garcia et al 1998) with slight modifications described by Calderon et al (2004). Amplicons were electrophoresed on agarose gels, digital images obtained, and the bands scored using GeneProfiler version 4.03 (Scanalytics, Merriefield, WA).…”