OBJECTIVE: To identify two closely related Brugia malayi and B. pahangi in cat reservoirs by using high resolution melting real-time PCR (HRM real-time PCR). MATERIAL AND METHOD: HRM analysis on the Corbett Rotor-Gene 6000 instrument was used to test 5 Brugia specimens by using five sets of specific primers for HhaI repetitive region (HR), small heat shock protein (SHP), small subunit ribosomal DNA (18S rDNA), internal transcribed spacer region (ITS), and trans-spliced leading Exon I gene (SLX1). RESULTS: HRM analysis of ITS and SLX clearly generated 2 profiles of B. malayi and B. pahangi while those of HR, 18S rDNA, and SHP could classify B. pahangi. CONCLUSION: HRM is a simple and rapid method for identification of two closely related B. malayi and B. pahangi in which it can detect both parasites within 30 min after real-time PCR detection. This assay is probe-free HRM and reduces a risk of PCR carryover. It does not require multiplex methods and DNA sequencing; therefore, HRM provides a new approach for genetic screening and rapid detection of closely related species in a clinical laboratory.
OBJECTIVE: To identify two closely related Brugia malayi and B. pahangi in cat reservoirs by using high resolution melting real-time PCR (HRM real-time PCR). MATERIAL AND METHOD: HRM analysis on the Corbett Rotor-Gene 6000 instrument was used to test 5 Brugia specimens by using five sets of specific primers for HhaI repetitive region (HR), small heat shock protein (SHP), small subunit ribosomal DNA (18S rDNA), internal transcribed spacer region (ITS), and trans-spliced leading Exon I gene (SLX1). RESULTS: HRM analysis of ITS and SLX clearly generated 2 profiles of B. malayi and B. pahangi while those of HR, 18S rDNA, and SHP could classify B. pahangi. CONCLUSION: HRM is a simple and rapid method for identification of two closely related B. malayi and B. pahangi in which it can detect both parasites within 30 min after real-time PCR detection. This assay is probe-free HRM and reduces a risk of PCR carryover. It does not require multiplex methods and DNA sequencing; therefore, HRM provides a new approach for genetic screening and rapid detection of closely related species in a clinical laboratory.
Authors: Ján Radvánský; Eva Bazsalovicsová; Ivica Králová-Hromadová; Gabriel Minárik; Ludevít Kádaši Journal: Parasitol Res Date: 2010-10-22 Impact factor: 2.289
Authors: Janina Demeler; Sabrina Ramünke; Sonja Wolken; Davide Ianiello; Laura Rinaldi; Jean Bosco Gahutu; Giuseppe Cringoli; Georg von Samson-Himmelstjerna; Jürgen Krücken Journal: PLoS One Date: 2013-04-19 Impact factor: 3.240
Authors: Hany Sady; Hesham M Al-Mekhlafi; Romano Ngui; Wahib M Atroosh; Ahmed K Al-Delaimy; Nabil A Nasr; Salwa Dawaki; Awatif M Abdulsalam; Init Ithoi; Yvonne A L Lim; Kek Heng Chua; Johari Surin Journal: Int J Mol Sci Date: 2015-07-16 Impact factor: 5.923