Advances in Real‐Time PCR: Application to Clinical Laboratory Diagnostics

Kaltenboeck, Bernhard; Wang, Chengming

doi:10.1016/s0065-2423(05)40006-2

Cited by 137 publications

(97 citation statements)

References 172 publications

(227 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Posteriormente se fueron incorporando distintas empresas como BioGene, Bioneer, Bio-Rad, Cepheid, Corbett Research, Idaho Technology, MJ Research, Roche, Applied Science, y Stratagene. En los equipos actuales, la detección del sistema incorpora módulos ópticos que son capaces de detectar secuencias seleccionadas marcadas con fluorocromos (6,10,11,13).…”

Section: Generalidadesunclassified

Bodas de plata de la reacción en cadena de la polimerasa (PCR)

Pinilla

Cubillos

Rodríguez³

2008

nova

View full text Add to dashboard Cite

ResumenInvenciones verdaderamente revolucionarias han promovido el cambio de pensamiento y la manera de trabajar en el ámbito del laboratorio. Una de estas invenciones es la reacción en cadena de la polimerasa, la cual ha aportado de manera significativa al conocimiento científico. Las diferentes metodologías que aplican la reacción en cadena de la polimerasa han permitido a los investigadores manipular la información genética de los organismos, facilitando procedimientos como la clonación y la secuenciación, entre otros, lo cual agilizó significativamente los resultados del Proyecto Genoma Humano. Existe diversidad de variantes de la reacción en cadena de la polimerasa convencional. Este escrito tiene por objetivo presentar una revisión sobre el tema, especialmente sobre la reacción en cadena de la polimerasa en tiempo real, debido a las ventajas que ofrece.Palabras clave: agentes intercalantes, cuantificación absoluta, cuantificación relativa, curvas de calibración, gen normalizador, sondas fluorogénicas. Abstract Silver anniversary of the Polymerase Chain Reaction (PCR)Truly revolutionary inventions have promoted the change of thought and the way to work in the laboratory. One of these inventions is the polymerase chain reaction (PCR), which has contributed significantly to the scientific knowledge. The different methodologies that use the polymerase chain reaction have allowed investigators to manipulate the genetic information of organisms, facilitating procedures like cloning and sequencing, among others, which sped the Human Genome Project results significantly. There are several variants of the conventional polymerase chain reaction. This work tries to present a revision on the subject, especially on the Real-time polymerase chain reaction, due to the advantages that it offers.

show abstract

Section: Generalidadesunclassified

Bodas de plata de la reacción en cadena de la polimerasa (PCR)

Pinilla

Cubillos

Rodríguez³

2008

nova

View full text Add to dashboard Cite

show abstract

“…According to the available information to date, Real Time PCR technique have been used for a number of tests to identify the pathogenic and saprophytic strain of Leptospira, which have been used for the diagnosis of Leptospirosis (12). Up to now, only a few tests are available for the differential diagnosis of pathogenic and saprophytic strains of Leptospira at the same time (13,14). The importance and necessity of early detection and diagnosis of pathogenic and saprophytic strains in human Leptospirosis is very obvious for treatment and epidemiologic studies.…”

Section: Introductionmentioning

confidence: 99%

Simultaneous Detection of Pathogenic and Saprophyte Leptospira in Human Plasma by Multiplex Taqman Real Time PCR

Alizadeh¹,

Javadi²,

Alizadeh³

et al. 2016

Biotech Health Sci

View full text Add to dashboard Cite

Background: Leptospirosis is an infectious disease caused by pathogenic and saprophytic Leptospira species. The clinical and laboratory diagnosis of this infection is complicated. However, timely diagnosis of leptospirosis is essential for treatment of this disease. Conventional laboratory methods are incapable in the early diagnosis of it. Molecular tests such as real time PCR are very efficient when diagnosing it.

show abstract

“…Real-time RT PCR is a convenient method for studying changes in gene expression, often used in biomedical research (7,8). However, its accuracy and reliability critically depend on the selection of a normalization strategy (9,10).…”

Section: Introductionmentioning

confidence: 99%

Hypoxanthine Guanine Phosphoribosyl Transferase Is the Most Stable Reference Gene for Gene Expression Analysis by Quantitative PCR in Peripheral Blood Mononuclear Cells from Women with the Polycystic Ovary Syndrome

Milutinović¹,

Macut²,

Antić³

et al. 2014

Journal of Medical Biochemistry

View full text Add to dashboard Cite

Summary Background: The polycystic ovary syndrome (PCOS) is a frequent endocrine disorder that affects women of reproductive age. As the syndrome is strongly associated with obesity, it is of interest to examine the gene expression diffe rences that accompany its development and the associ a ted metabolic disturbances. Real-time RT PCR is a standard method for studying changes in gene expression. However, to obtain accurate and reliable results, validation of reference genes is obligatory. The aim of this study was to identify a suitable reference for the normalization of gene expression in peripheral blood mononuclear cells (PBMCs) from obese and normal-weight women with PCOS. Methods: The expression stability of four potential reference genes: hypoxanthine guanine phosphoribosyl trans-ferase 1 (HPRT), β-actin (BA), β2-microglobulin (B2M) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), was assessed in PBMCs from healthy women, and from normal-weight and obese women with PCOS. The variability in the expression of potential reference genes was analyzed by the TaqMan real-time RT PCR method, using GeNorm and NormFinder software packages. Results: Direct comparison of cycle threshold (Ct) values showed inter-individual variations for all validated genes, the Ct values of HPRT being less variable than those of BA, GAPDH and B2M. Both software packages pointed to HPRT as the most steadily expressed gene in the PBMCs of women with PCOS and healthy controls. Conclusions: Cross-validation of the expression stability of four potential reference genes identified HPRT as the most stable reference, suitable for further investigations of gene expression in PBMCs from women with PCOS.

show abstract

Advances in Real‐Time PCR: Application to Clinical Laboratory Diagnostics

Cited by 137 publications

References 172 publications

Bodas de plata de la reacción en cadena de la polimerasa (PCR)

Bodas de plata de la reacción en cadena de la polimerasa (PCR)

Simultaneous Detection of Pathogenic and Saprophyte Leptospira in Human Plasma by Multiplex Taqman Real Time PCR

Hypoxanthine Guanine Phosphoribosyl Transferase Is the Most Stable Reference Gene for Gene Expression Analysis by Quantitative PCR in Peripheral Blood Mononuclear Cells from Women with the Polycystic Ovary Syndrome

Contact Info

Product

Resources

About