A novel quantitative multiplex NASBA method: application to measuring tissue factor and CD14 mRNA levels in human monocytes

Deursen, P. B. H. van; Gunther, A. W.; Riel, Caroline C. Spaargaren-van; Eijnden, Mark M. E. D. van den; Vos, Hans L.; Gemen, Bob van; Strijp, Dianne A. M. W. van; Tacken, N.; Bertina, Rogier M.

doi:10.1093/nar/27.17.e15

Cited by 23 publications

(7 citation statements)

References 23 publications

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“…The template DNA was removed by treatment with DNase I (Takara Shuzou, Kyoto, Japan), after which the resultant genomic RNA fragments were quantified by spectroscopy and the number of molecules calculated from the optical density. DNase treatment and RNA quantification were essentially as described in van Deursen et al. (1999).…”

Section: Methodsmentioning

confidence: 99%

“…Unlike PCR-based methods, however, the presence of amplicons cannot be detected by conventional agarose electrophoresis, because a formaldehydedenaturing system (Sambrook et al 1989) is needed for the precise sizing of single-stranded RNA. Therefore, a sandwich hybridization method has been used for postamplification analysis of NASBA (Guatelli et al 1990;van Deursen et al 1999). Applications of homogeneous, fluorescence-based assays for the NASBA method have also been developed.…”

Section: Introductionmentioning

confidence: 99%

“…Therefore, a sandwich hybridization method has been used for postamplification analysis of NASBA (Guatelli et al. 1990; van Deursen et al. 1999).…”

Section: Introductionmentioning

confidence: 99%

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Real-time nucleic acid sequence-based amplification (NASBA) using an adenine-induced quenching probe and an intercalator dye

Kouguchi¹,

Teramoto²,

Kuramoto³

2010

Journal of Applied Microbiology

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Aims: We found that an adenine base caused fluorescence quenching of a fluorescein (FL)‐labelled probe in DNA:RNA hybrid sequences, and applied this finding to a nucleic acid sequence–based amplification (NASBA) method. Methods and Results: The present NASBA method employed a probe containing an FL‐modified thymine at its 3′ end and ethidium bromide (EtBr) on the basis of a combination of adenine‐induced quenching and fluorescence resonance energy transfer (FRET) between the FL donor and EtBr acceptor. This NASBA was used to detect Shiga toxin (STX) stx‐specific mRNA in STX‐producing Escherichia coli, demonstrating rapid quantification of the target gene with high sensitivity. Conclusion: Although the inherent quenching effect of adenine was inferior to that of guanine, FRET between the FL and EtBr moieties enhanced the adenine‐induced quenching, allowing rapid and sensitive real‐time NASBA detection. Significance and Impact of the Study: This study gives a novel real‐time diagnostic system based on NASBA for a sensitive mRNA (or viral RNA) detection.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Real-time nucleic acid sequence-based amplification (NASBA) using an adenine-induced quenching probe and an intercalator dye

Kouguchi¹,

Teramoto²,

Kuramoto³

2010

Journal of Applied Microbiology

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“…Recently, real-time NASBA was used for the detection of IAV H1N1 and H3N2 subtypes, influenza B virus, respiratory syncytial virus (RSV), and metapneumovirus isolated from the swabs of patients with respiratory tract infections [ 100 ]. Multiplex NASBA is an improved method, which is suitable for the simultaneous quantification of multiple target genes [ 101 ]. Multiplex NASBA with an enzyme-linked oligonucleotide capture optical detection method can distinguish between a group of viruses causing common lower respiratory tract infections, including influenza A and B viruses, human parainfluenza virus 1-4, RSV, rubella virus, and Coxsackie virus [ 102 ].…”

Section: Dna Amplification Under Isothermal Conditionsmentioning

confidence: 99%

Updated Values for Molecular Diagnosis for Highly Pathogenic Avian Influenza Virus

Sakurai¹,

Shibasaki²

2012

Viruses

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Highly pathogenic avian influenza (HPAI) viruses of the H5N1 strain pose a pandemic threat. H5N1 strain virus is extremely lethal and contagious for poultry. Even though mortality is 59% in infected humans, these viruses do not spread efficiently between humans. In 1997, an outbreak of H5N1 strain with human cases occurred in Hong Kong. This event highlighted the need for rapid identification and subtyping of influenza A viruses (IAV), not only to facilitate surveillance of the pandemic potential of avian IAV, but also to improve the control and treatment of infected patients. Molecular diagnosis has played a key role in the detection and typing of IAV in recent years, spurred by rapid advances in technologies for detection and characterization of viral RNAs and proteins. Such technologies, which include immunochromatography, quantitative real-time PCR, super high-speed real-time PCR, and isothermal DNA amplification, are expected to contribute to faster and easier diagnosis and typing of IAV.

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“…[26], Aspergillus spp. [27], Plasmodium falciparum [28], macrophage-derived chemokine mRNA [29], tissue factor mRNA [30], and human TNF-a mRNA [31], among many others. Other advantages of NASBA include the fact that amplification levels can reach 10 9 -10 12 after 2 h. In addition, the amplification process is isothermal (41°C) and does not require a thermal cycler or other specialised amplification equipment.…”

mentioning

confidence: 99%

A method to detect major serotypes of foot-and-mouth disease virus

Collins

Fung

et al. 2002

Biochemical and Biophysical Research Communications

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Nucleic acid sequence-based amplification (NASBA) is an isothermal technique that allows the rapid amplification of specific regions of nucleic acid obtained from a diverse range of sources. It is especially suitable for amplifying RNA sequences. A rapid and specific NASBA technique was developed, allowing the detection of foot-and-mouth disease virus genetic material in a range of sample material, including preserved skin biopsy material from infected animals, vaccines prepared from denatured cell-free material, and cell-free antigen-based detection kits. A single pair of DNA oligonucleotide primers was able to amplify examples of all major FMD virus subtypes. The amplified viral RNA was detected by electrochemiluminescence. The method was at least as sensitive as existing cell-free antigen detection methods.

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A novel quantitative multiplex NASBA method: application to measuring tissue factor and CD14 mRNA levels in human monocytes

Cited by 23 publications

References 23 publications

Real-time nucleic acid sequence-based amplification (NASBA) using an adenine-induced quenching probe and an intercalator dye

Real-time nucleic acid sequence-based amplification (NASBA) using an adenine-induced quenching probe and an intercalator dye

Updated Values for Molecular Diagnosis for Highly Pathogenic Avian Influenza Virus

A method to detect major serotypes of foot-and-mouth disease virus

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