Rapid Detection and Identification of Wuchereria bancrofti, Brugia malayi, B. pahangi, and Dirofilaria immitis in Mosquito Vectors and Blood Samples by High Resolution Melting Real-Time PCR
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
A simple, rapid, and high-throughput method for detection and identification of Wuchereria bancrofti, Brugia malayi, Brugia pahangi, and Dirofilaria immitis in mosquito vectors and blood samples was developed using a real-time PCR combined with high-resolution melting (HRM) analysis. Amplicons of the 4 filarial species were generated from 5S rRNA and spliced leader sequences by the real-time PCR and their melting temperatures were determined by the HRM method. Melting of amplicons from W. bancrofti, B. malayi, D. immitis, and B. pahangi peaked at 81.5±0.2℃, 79.0±0.3℃, 76.8±0.1℃, and 79.9±0.1℃, respectively. This assay is relatively cheap since it does not require synthesis of hybridization probes. Its sensitivity and specificity were 100%. It is a rapid and technically simple approach, and an important tool for population surveys as well as molecular xenomonitoring of parasites in vectors.
Abstract
A simple, rapid, and high-throughput method for detection and identification of Wuchereria bancrofti, Brugia malayi, Brugia pahangi, and Dirofilaria immitis in mosquito vectors and blood samples was developed using a real-time PCR combined with high-resolution melting (HRM) analysis. Amplicons of the 4 filarial species were generated from 5S rRNA and spliced leader sequences by the real-time PCR and their melting temperatures were determined by the HRM method. Melting of amplicons from W. bancrofti, B. malayi, D. immitis, and B. pahangi peaked at 81.5±0.2℃, 79.0±0.3℃, 76.8±0.1℃, and 79.9±0.1℃, respectively. This assay is relatively cheap since it does not require synthesis of hybridization probes. Its sensitivity and specificity were 100%. It is a rapid and technically simple approach, and an important tool for population surveys as well as molecular xenomonitoring of parasites in vectors.
참고문헌 (28)
- Weil GJ, "Diagnostic tools for filariasis elimination programs", Trends Parasitol, 23, pp.78~82 (2007)
- World Health Organization, "Lymphatic filariasis: the disease and its control. Fifth report of the WHO Expert Committee on Filariasis", Tech Rep Ser, 821, pp.1~71 (1992)
- Triteeraprapab S, "High prevalence of Wuchereria bancrofti infection among Myanmar migrants in Thailand", Ann Trop Med Parasitol, 95, pp.535~538 (2001)
- Center for Disease Control and Prevention, "Recommendation of International Task Force for Disease Eradication. Morbidity and Mortality Weekly Report", Recommendation and Reports, 42, pp.1~38 (1993)
- Orihel TC, "Morphology of the larval stages of Dirofilaria immitis in the dog", J Parasitol, 47, pp.251~262 (1961)
- Denham DA, "Brugian filariasis : epidemiological and experimental studies", Adv Parasitol, 15, pp.243~309 (1977)
- Lim KC, "Human Ecology and Infectious Disease", pp.49~72 (1983), Academic Press
- Palmieri JR, "Evidence of possible natural infections of man with Brugia pahangi in South Kalimantan(Borneo), Indonesia", Trop Geogr Med, 37, pp.239~244 (1985)
- Simón F, "Human and animal dirofilariasis : the emergence of a zoonotic mosaic", Clin Microbiol Rev, 25, pp.507~544 (2012)
- Latrofa MS, "A multiplex PCR for the simultaneous detection of species of filarioids infesting dogs", Acta Trop, 122, pp.150~154 (2012)
- Nuchprayoon S, "Differentiation of Brugia malayi and Brugia pahangi by PCR-RFLP of ITS-1 and ITS-2", Southeast Asian J Trop Med Public Health, 34, pp.67~73 (2003)
- Lulitanond V, "Rapid detection of Wuchereria bancrofti in mosquitoes by LightCycler polymerase chain reaction and melting curve analysis", Parasitol Res, 94, pp.337~341 (2004)
- Thanchomnang T, "Rapid detection of Brugia malayi in mosquito vectors using a real-time fluorescence resonance energy transfer PCR and melting curve analysis", Am J Trop Med Hyg, 78, pp.509~513 (2008)
- Thanchomnang T, "Differential detection of Brugia malayi and Brugia pahangi by real-time fluorescence resonance energy transfer PCR and its evaluation for diagnosis of B. pahangi-infected dogs", Parasitol Res, 106, pp.621~625 (2010)
- Thanchomnang T, "Rapid detection of Dirofilaria immitis in mosquito vectors and dogs using a real-time fluorescence resonance energy transfer PCR and melting curve analysis", Vet Parasitol, 168, pp.255~260 (2010)
- Duscher G, "Detection of Dirofilaria in Austrian dogs", Berl Munch Tierarztl Wochenschr, 122, pp.199~203 (2009)
- Wongkamchai S, "Rapid detection and identification of Brugia malayi, B. pahangi, and Dirofilaria immitis by high-resolution melting assay", Vector Borne Zoonotic Dis, 13, pp.31~36 (2013)
- Wittwer CT, "High-resolution genotyping by amplicon melting analysis using LCGreen", Clin Chem, 49, pp.853~860 (2003)
- Choochote W, "Aedes(Finlaya)togoi Theobald 1907, Chanthaburi strain, a laboratory vector in studies of filariasis in Thailand", Southeast Asian J Trop Med Public Health, 18, pp.259~260 (1987)
- Maleewong W, "Scanning electron microscopic study of third-stage larva of Wuchereria bancrofti and Brugia malayi in Thailand", Southeast Asian J Trop Med Public Health, 18, pp.261~264 (1987)
- Choochote W, "Aedes albolateralis, a potential vector of nocturnally subperiodic Wuchereria bancrofti and dengue type 2 virus", Southeast Asian J Trop Med Public Health, 32, pp.621~624 (2001)
- World Health Organization., "Basic laboratory methods in medical parasitology", World Health Organization, 39~61 (1991)
- Yen PK, "Histochemical differentiation of Brugia, Wuchereria, Dirofilaria and Breinlia microfilariae", Ann Trop Med Parasitol, 72, pp.157~162 (1978)
- Chungpivat S, "The differentiation of microfilariae in dogs and cats using Giemsa’s staining and the detection of acid phosphatase activity", J Thai Vet Pract, 20, pp.47~55 (2008)
- Areekit S, "High resolution melting real-time PCR for rapid discrimination between Brugia malayi and Brugia pahangi", J Med Assoc Thai, 92(suppl 3), pp.S24~S28 (2009)
- World Health Organization, "Global programme to eliminate lymphatic filariasis", Wkly Epidemiol Rec, 82, pp.361~380 (2007)
- Ririe KM, "Product differentiation by analysis of DNA melting curves during the polymerase chain reaction", Anal Biochem, 245, pp.154~160 (1997)
- Jariya P, "Dirofilaria repens from the eyelid of a woman in Thailand", Am J Trop Med Hyg, 32, pp.1456~1457 (1983)
