In Vitro Nucleic Acid Amplification Techniques

Datta, Vivekanand; Hayden, Randall T.

doi:10.1128/9781555816834.ch3

Cited by 4 publications

(4 citation statements)

References 222 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A reverse transcription PCR step is performed prior to the first PCR cycle. Amplification of target nucleic acids is then accomplished using the principle of nested PCR which uses a two stages approach to increase the sensitivity and specificity of the PCR reaction [64].…”

Section: The Filmarray Instrumentmentioning

confidence: 99%

“…Multiplex molecular diagnostic assays are ideal for infections with a wide differential diagnosis. However, the design of broadly multiplexed assays is challenging as multiplex PCR assays are subject to decreased sensitivity due in part to, targets' competition and primer dimers formation [64]. The overall sensitivity and specificity of the FilmArray RP in published reports varied between 85-100% and 100%, respectively with the sensitivity of the adenovirus assays remaining a challenge even in the new version of the assay.…”

Section: Expert Commentarymentioning

confidence: 99%

See 1 more Smart Citation

The FilmArray® respiratory panel: an automated, broadly multiplexed molecular test for the rapid and accurate detection of respiratory pathogens

Babady

2013

Expert Review of Molecular Diagnostics

109

View full text Add to dashboard Cite

The FilmArray Respiratory Panel (RP) (BioFire(™) Diagnostics, Inc., Salt Lake City, UT, USA) is the first multiplex molecular panel cleared by the US FDA for the detection of both bacterial and viral respiratory pathogens in nasopharygeal swabs. The FilmArray RP targets 20 pathogens including 17 viruses and subtypes and three bacteria, and is performed with minimal sample manipulation. The FilmArray RP has a fully automated sample-to-answer workflow with a turn-around-time of approximately 1 h. The reported sensitivity and specificity of the assay ranges from 80 to 100 and 100%, respectively, with the sensitivity for the adenovirus as low as 46%. A new version of the FilmArray RP assay (version 1.7) with improved sensitivity for the adenovirus was released in 2013. The performance characteristics and simplified workflow have allowed its implementation in a wide range of laboratories. The FilmArray RP has changed the diagnostic landscape and will have a significant impact on the care of patients with respiratory tract infection.

show abstract

Section: The Filmarray Instrumentmentioning

confidence: 99%

Section: Expert Commentarymentioning

confidence: 99%

The FilmArray® respiratory panel: an automated, broadly multiplexed molecular test for the rapid and accurate detection of respiratory pathogens

Babady

2013

Expert Review of Molecular Diagnostics

109

View full text Add to dashboard Cite

show abstract

“…Design and optimization of micro-heater electrodes for PCR thermal cycling application.-Temperature control is essential for an integrated PCR micro-device since performance of PCR reaction is greatly impacted by the temperature set-points and temperature ramp rates during thermal cycling. 7,10 Poor temperature control can result in low PCR efficiency and non-specific probe-target DNA binding and amplification. Methods for chip-based temperature control can be classified as: contact or non-contact.…”

Section: Theorymentioning

confidence: 99%

“…Nowadays, this is almost always done through a "one tube" technique, involving a reaction mixture containing both a reverse transcriptase enzyme and a thermostable DNA polymerase, with the two enzymatic reactions performed serially through temperature control. 7 z E-mail: kaler@ucalgary.ca For influenza A testing, ProvLab has adapted an assay designed and implemented by the Centers for Disease Control, as described in, 8 and which uses the common methodology of hydrolysis probe for detection. 6 For influenza C, the real time RT-PCR method developed at ProvLab, 2 was in fact validated to work with either a hydrolysis probe or a beacon probe.…”

mentioning

confidence: 99%

Droplet Microfluidic Chip Based Nucleic Acid Amplification and Real-Time Detection of Influenza Viruses

Prakash

Wong

et al. 2013

J. Electrochem. Soc.

View full text Add to dashboard Cite

Miniaturized bio-diagnostic devices have the potential to allow for rapid pathogen screening in clinical patient samples, as a low cost and portable alternative to conventional bench-top equipment. Miniaturization of key bio-diagnostic techniques, such as: nucleic acid detection and quantification, polymerase chain reaction (PCR), DNA fingerprinting, enzyme linked immunosorbent assay (ELISA), results in substantial reduction of reaction volumes (expensive samples/reagents) and shorter reaction times. Droplet microfluidics (DMF) is one of several miniaturized bio-sample handling techniques available for manipulating clinical samples and reagents in microliter (10 −6 L) to picoliter (10 −12 L) volume regime. Electro-actuation of sample and reagent in the form of droplets in the aforementioned volume regime, using dielectrophoresis (DEP) and/or Electrowetting (EW) are achieved by means of patterned, insulated metal electrodes on one or more substrates. In this work, we have utilized electro-actuation based DMF technology, integrated with suitably tailored resistive micro-heaters and temperature sensors, to achieve chip based real-time, quantitative PCR (qRT-PCR). This qRT-PCR micro-device was utilized to detect and quantify the presence of influenza A and C virus nucleic acids, using in-vitro synthesized viral RNA segments. The experimental analysis of the DMF micro-device confirms its capabilities in qRT-PCR based detection and quantification of pathogen samples, with accuracy levels comparable to established commercial bench-top equipment (PCR efficiency ∼95%). The limit of detection (LOD) of the chip based qRT-PCR technique was estimated to be ∼5 copies of template RNA per PCR reaction.

show abstract

Impacts and Challenges of Advanced Diagnostic Assays for Transplant Infectious Diseases

Babady

Lee

Papanicolaou

et al. 2019

Principles and Practice of Transplant Infectious Diseases

View full text Add to dashboard Cite

In Vitro Nucleic Acid Amplification Techniques

Cited by 4 publications

References 222 publications

The FilmArray® respiratory panel: an automated, broadly multiplexed molecular test for the rapid and accurate detection of respiratory pathogens

The FilmArray® respiratory panel: an automated, broadly multiplexed molecular test for the rapid and accurate detection of respiratory pathogens

Droplet Microfluidic Chip Based Nucleic Acid Amplification and Real-Time Detection of Influenza Viruses

Impacts and Challenges of Advanced Diagnostic Assays for Transplant Infectious Diseases

Contact Info

Product

Resources

About