BACKGROUND AND OBJECTIVE: We describe here a nanodiagnostic colorimetric assay for 18S rRNA of Leishmania pathogens that uses nucleic acid sequence-based amplification (NASBA) and gold nanorods (GNRs). METHODS: NASBA primers targeting 18S rRNA were used for amplification of RNA in an isothermal process. The electrostatic interactions between the phosphate groups of the RNA amplicons and the cetyl trimethylammonium bromide layer on the GNRs resulting in their aggregation. This phenomenon changes the color of the GNR solution from red to purple. RESULTS: Our data showed 100% sensitivity and 80% specificity with the colorimetric assay compared with results using reverse transcription polymerase chain reaction. CONCLUSION: The nanodiagnostic method we describe simplifies the detection of NASBA amplicons without the need for gel electrophoresis.
BACKGROUND AND OBJECTIVE: We describe here a nanodiagnostic colorimetric assay for 18S rRNA of Leishmania pathogens that uses nucleic acid sequence-based amplification (NASBA) and gold nanorods (GNRs). METHODS: NASBA primers targeting 18S rRNA were used for amplification of RNA in an isothermal process. The electrostatic interactions between the phosphate groups of the RNA amplicons and the cetyl trimethylammonium bromide layer on the GNRs resulting in their aggregation. This phenomenon changes the color of the GNR solution from red to purple. RESULTS: Our data showed 100% sensitivity and 80% specificity with the colorimetric assay compared with results using reverse transcription polymerase chain reaction. CONCLUSION: The nanodiagnostic method we describe simplifies the detection of NASBA amplicons without the need for gel electrophoresis.
Authors: Wendy F van der Meide; Gerard J Schoone; William R Faber; Jim E Zeegelaar; Henry J C de Vries; Yusuf Ozbel; Rudy F M Lai A Fat; Leíla I A R C Coelho; Masoom Kassi; Henk D F H Schallig Journal: J Clin Microbiol Date: 2005-11 Impact factor: 5.948
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Authors: B van Gemen; R van Beuningen; A Nabbe; D van Strijp; S Jurriaans; P Lens; T Kievits Journal: J Virol Methods Date: 1994-09 Impact factor: 2.014
Authors: S A Omar; P F Mens; G J Schoone; A Yusuf; J Mwangi; S Kaniaru; G A A Omer; H D F H Schallig Journal: Exp Parasitol Date: 2005-05 Impact factor: 2.011
Authors: A Schubach; F Haddad; M P Oliveira-Neto; W Degrave; C Pirmez; G Grimaldi; O Fernandes Journal: J Infect Dis Date: 1998-09 Impact factor: 5.226