title: Rapid detection and identification of Brugia malayi, B. pahangi, and Dirofilaria immitis by high-resolution melting assay.
abstract: Human lymphatic filariasis is caused by filarial worms such as Brugia malayi for which the major reservoir is domestic cats. However, domestic cats or dogs also carry nonhuman filaria such as Brugia pahangi and Dirofilaria immitis. We have developed a single-tube, real-time PCR with a high-resolution melting (HRM) analysis assay for detection and identification of B. malayi, B. pahangi, and D. immitis in blood samples. The designated primer pair in the PCR can amplify a 114-bp region of mitochondrial 12S rRNA genes of these filarial worms. Subsequently, the HRM assay showed a specific melting temperature for each species. The assay showed the highest sensitivity and specificity in comparison with DNA sequences after assessment with 34 cat and 14 dog blood samples. This assay could be helpful for epidemiological studies of reservoirs and vectors.
authors.affiliation: Department of Parasitology, Faculty of Medicine Siriraj Hospital, Bangkok, Thailand.
authors.forename: Sirichit
authors.initials: S
authors.lastname: Wongkamchai
authors.forename: Nuntiya
authors.initials: N
authors.lastname: Monkong
authors.forename: Pakpimom
authors.initials: P
authors.lastname: Mahannol
authors.forename: Piyanan
authors.initials: P
authors.lastname: Taweethavonsawat
authors.forename: Sumat
authors.initials: S
authors.lastname: Loymak
authors.forename: Suporn
authors.initials: S
authors.lastname: Foongladda
mesh_terms: D000818:Animals; D001483:Base Sequence; D017178:Brugia malayi / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D017179:Brugia pahangi / Q000235:genetics / Q000302:isolation & purification*; D002415:Cats; D017931:DNA Primers / Q000235:genetics; D004272:DNA, Mitochondrial / Q000097:blood / Q000235:genetics; D016054:DNA, Protozoan / Q000097:blood / Q000235:genetics; D004275:DNA, Ribosomal / Q000097:blood / Q000235:genetics; D004183:Dirofilaria immitis / Q000235:genetics / Q000302:isolation & purification*; D004184:Dirofilariasis / Q000453:epidemiology / Q000469:parasitology; D004197:Disease Reservoirs; D004285:Dogs; D005368:Filariasis / Q000453:epidemiology / Q000469:parasitology*; D006801:Humans; D008969:Molecular Sequence Data; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D013785:Thailand / Q000453:epidemiology; D044366:Transition Temperature
chemical_list: D017931:DNA Primers; D004272:DNA, Mitochondrial; D016054:DNA, Protozoan; D004275:DNA, Ribosomal