Warning: Undefined array key "mm" in /www/wwwroot/www.ai-bt.com/si.php on line 10 Deprecated: trim(): Passing null to parameter #1 ($string) of type string is deprecated in /www/wwwroot/www.ai-bt.com/si.php on line 10 Quantitative detection and differentiation of free-living amoeba species using SYBR green-based real-time PCR melting curve analysis.

Literature DB >> 17106802

Quantitative detection and differentiation of free-living amoeba species using SYBR green-based real-time PCR melting curve analysis.

Jonas Behets¹, Priscilla Declerck, Yasmine Delaedt, Lieve Verelst, Frans Ollevier.

Abstract

Real-time polymerase chain reaction melting curve analysis (MCA) allows differentiation of several free-living amoebae species. Distinctive characteristics were found for Naegleria fowleri, N. lovaniensis, N. australiensis, N. gruberi, Hartmanella vermiformis, and Willaertia magna. Species specificity of the amplicons was confirmed using agarose gel electrophoresis and sequence-based approaches. Amplification efficiency ranged from 91% to 98%, indicating the quantitative potential of the assay. This MCA approach can be used for quantitative detection of free-living amoebae after cultivation but also as a culture-independent detection method.

Entities: Chemical Species

Mesh：

Year: 2006 PMID： 17106802 DOI： 10.1007/s00284-006-0241-0

Source DB: PubMed Journal: Curr Microbiol ISSN： 0343-8651 Impact factor: 2.188

22 in total

Review 1. Genome structure and evolution of Naegleria and its relatives.

Authors: C G Clark
Journal: J Protozool Date: 1990 Jul-Aug

2. Comparison of SYTO9 and SYBR Green I for real-time polymerase chain reaction and investigation of the effect of dye concentration on amplification and DNA melting curve analysis.

Authors: Paul T Monis; Steven Giglio; Christopher P Saint
Journal: Anal Biochem Date: 2005-05-01 Impact factor: 3.365

Review 3. Genomic and evolutionary aspects of Mimivirus.

Authors: M Suzan-Monti; B La Scola; D Raoult
Journal: Virus Res Date: 2005-09-21 Impact factor: 3.303

Review 4. Emerging technologies for the detection and genetic characterization of protozoan parasites.

Authors: Paul T Monis; Steven Giglio; Alexandra R Keegan; R C Andrew Thompson
Journal: Trends Parasitol Date: 2005-07

5. Development of a real-time PCR assay for quantification of Acanthamoeba trophozoites and cysts.

Authors: Delphine Rivière; Florence Ménard Szczebara; Jean-Marc Berjeaud; Jacques Frère; Yann Héchard
Journal: J Microbiol Methods Date: 2005-05-31 Impact factor: 2.363

6. Legionella confirmation using real-time PCR and SYTO9 is an alternative to current methodology.

Authors: Steven Giglio; Paul T Monis; Christopher P Saint
Journal: Appl Environ Microbiol Date: 2005-12 Impact factor: 4.792

7. Use of multiplex PCR and PCR restriction enzyme analysis for detection and exploration of the variability in the free-living amoeba Naegleria in the environment.

Authors: Michel Pélandakis; Pierre Pernin
Journal: Appl Environ Microbiol Date: 2002-04 Impact factor: 4.792

Quantitative detection and differentiation of free-living amoeba species using SYBR green-based real-time PCR melting curve analysis.

Review 1. Genome structure and evolution of Naegleria and its relatives.

2. Comparison of SYTO9 and SYBR Green I for real-time polymerase chain reaction and investigation of the effect of dye concentration on amplification and DNA melting curve analysis.

Review 3. Genomic and evolutionary aspects of Mimivirus.

Review 4. Emerging technologies for the detection and genetic characterization of protozoan parasites.

5. Development of a real-time PCR assay for quantification of Acanthamoeba trophozoites and cysts.

6. Legionella confirmation using real-time PCR and SYTO9 is an alternative to current methodology.

7. Use of multiplex PCR and PCR restriction enzyme analysis for detection and exploration of the variability in the free-living amoeba Naegleria in the environment.

8. Detection and genotyping of oocysts of Cryptosporidium parvum by real-time PCR and melting curve analysis.

9. Design and evaluation of a specific primer pair for the diagnosis and identification of Acanthamoeba polyphaga.

10. Detection of Naegleria spp. and Naegleria fowleri: a comparison of flagellation tests, ELISA and PCR.

1. Comparison of real-time PCR methods for the detection of Naegleria fowleri in surface water and sediment.

Review 2. Methodological approaches for monitoring opportunistic pathogens in premise plumbing: A review.