SYBR Green-based real-time PCR assay for detection of VKORC1 and CYP2C9 polymorphisms that modulate warfarin dose requirement

Abstract Trichomonas gallinae are parasitic flagellates of importance in wild and domestic birds. The parasite is worldwide distributed, and Columbine birds are its main host. Current research focuses mostly on epidemiological and phylogenetic studies. However, there is still a lack of knowledge regarding parasite-host interaction or therapy development. Real-time PCR is a useful tool for diagnostic and quantification of gene copies in a determined sample. By amplification of a 113-bp region of the 18S small subunit ribosomal RNA gene, a SYBR green-based real-time PCR assay was developed. A standard curve was prepared for quantification analysis. Assay efficiency, linearity, and dissociation analysis were successfully performed. Specificity, sensibility, and reproducibility analysis were tested. This assay could be a useful tool not only for diagnostic purposes but also for future in vivo and in vitro T. gallinae studies.

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Development of SYBR Green I-based real-time PCR assay for detection of drug resistance mutations in cytomegalovirus

Journal of Virological Methods ◽

10.1016/j.jviromet.2007.12.011 ◽

2008 ◽

Vol 149 (1) ◽

pp. 129-135 ◽

Cited By ~ 7

Author(s):

Jing-bo Liu ◽

Zheng Zhang

Keyword(s):

Drug Resistance ◽

Real Time ◽

Real Time Pcr ◽

Sybr Green ◽

Sybr Green I ◽

Resistance Mutations ◽

Pcr Assay ◽

Drug Resistance Mutations

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Detection and species identification of microsporidial infections using SYBR Green real-time PCR

Journal of Medical Microbiology ◽

10.1099/jmm.0.026781-0 ◽

2011 ◽

Vol 60 (4) ◽

pp. 459-466 ◽

Cited By ~ 29

Author(s):

Spencer D. Polley ◽

Samuel Boadi ◽

Julie Watson ◽

Alan Curry ◽

Peter L. Chiodini

Keyword(s):

Real Time ◽

Species Identification ◽

Real Time Pcr ◽

Simultaneous Detection ◽

Enterocytozoon Bieneusi ◽

Sybr Green ◽

Pcr Assay ◽

Real Time Analysis ◽

Highly Sensitive ◽

Stool Samples

Diagnosis of microsporidial infections is routinely performed by light microscopy, with unequivocal non-molecular species identification achievable only through electron microscopy. This study describes a single SYBR Green real-time PCR assay for the simultaneous detection and species identification of such infections. This assay was highly sensitive, routinely detecting infections containing 400 parasites (g stool sample)−1, whilst species identification was achieved by differential melt curves on a Corbett Life Science Rotor-Gene 3000. A modification of the QIAamp DNA tissue extraction protocol allowed the semi-automated extraction of DNA from stools for the routine diagnosis of microsporidial infection by real-time PCR. Of 168 stool samples routinely analysed for microsporidian spores, only five were positive by microscopy. By comparison, 17 were positive for microsporidial DNA by real-time analysis, comprising 14 Enterocytozoon bieneusi, one Encephalitozoon cuniculi and two separate Pleistophora species infections.

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Molecular identification of Panthera Leo materials by SYBR green I real-time PCR assay

Journal of Biotechnology ◽

10.1016/j.jbiotec.2010.08.325 ◽

2010 ◽

Vol 150 ◽

pp. 125-125

Author(s):

Junyi Xu ◽

Jijuan Cao ◽

Qiuyan Wang

Keyword(s):

Real Time ◽

Molecular Identification ◽

Real Time Pcr ◽

Sybr Green ◽

Sybr Green I ◽

Panthera Leo ◽

Pcr Assay

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Quantitative Detection of Listeria monocytogenes in Biofilms by Real-Time PCR

Applied and Environmental Microbiology ◽

10.1128/aem.71.4.2190-2194.2005 ◽

2005 ◽

Vol 71 (4) ◽

pp. 2190-2194 ◽

Cited By ~ 42

Author(s):

Morgan Guilbaud ◽

Pierre de Coppet ◽

Fabrice Bourion ◽

Cinta Rachman ◽

Hervé Prévost ◽

...

Keyword(s):

Listeria Monocytogenes ◽

Real Time ◽

Real Time Pcr ◽

Quantitative Method ◽

Sybr Green ◽

Sybr Green I ◽

Quantitative Detection ◽

Pcr Assay

ABSTRACT A quantitative method based on a real-time PCR assay to enumerate Listeria monocytogenes in biofilms was developed. The specificity for L. monocytogenes of primers targeting the listeriolysin gene was demonstrated using a SYBR Green I real-time PCR assay. The number of L. monocytogenes detected growing in biofilms was 6 × 102 CFU/cm2.

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