A netlike rolling circle nucleic acid amplification technique

The Analyst ◽  
2015 ◽  
Vol 140 (1) ◽  
pp. 74-78 ◽  
Author(s):  
Xiaoli Zhu ◽  
Chang Feng ◽  
Bin Zhang ◽  
Hui Tong ◽  
Tao Gao ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Rolling Circle Amplification ◽  
Nucleic Acid Amplification ◽  
Amplification Efficiency ◽  
Rolling Circle ◽  
Isothermal Nucleic Acid Amplification ◽  
Amplification Technique ◽  
Nucleic Acid Amplification Technique ◽  
Network Morphology

An isothermal nucleic acid amplification technique termed as netlike rolling circle amplification is proposed. Dense and uniform network morphology of amplified products is first observed, suggesting the ultrahigh amplification efficiency.

scholarly journals Strategies for Isothermal Amplification of Nucleic Acids: are they Ready to be Applied in Point of Care Diagnosis of Mycosis?

2020 ◽  
Vol 11 (3) ◽  
pp. 10559-10571
Keyword(s):  
Nucleic Acid ◽  
Clinical Laboratory ◽  
Multiple Displacement Amplification ◽  
Rolling Circle Amplification ◽  
High Sensitivity ◽  
Isothermal Amplification ◽  
Nucleic Acid Amplification ◽  
Clinical Samples ◽  
Rolling Circle ◽  
Isothermal Nucleic Acid Amplification

The early detection of invasive fungal infection (IFD) is significant in order to decrease mortality in susceptible patients. There is, therefore, a need for sensitive and specific fungal species detection assays in a clinical laboratory for early targeted therapy. The isothermal amplification method may be useful for the screening of fungal isolates, especially in resource-poor settings. Therefore, our aim was to review the isothermal nucleic acid amplification methods and their applications in fungal pathogen detection. Out of 50 reported studies, 28, 12, 6, 2, and 2 studies used the isothermal-based assays of a loop-mediated isothermal amplification (LAMP), nucleic acid sequence-based amplification (NASBA), rolling circle amplification (RCA), multiple displacement amplification (MDA) and polymerase Spiral Reaction (PSR), respectively. Thirty-two studies used clinical samples, 18 pure culture, and four environmental samples. The diagnostic accuracy of isothermal nucleic acid amplification testing for pathogenic fungal was reported as high (sensitivity 0.89–1.0 and specificity 0.63–1.0) in all studies irrespective of the sample tested. Although the isothermal-based assays showed high sensitivity and specificity in reported studies, it is still poorer than that of PCR assays. However, improving the assay to make it simpler, more effective, and inexpensive compared with newer PCR methods are still needed.

scholarly journals Risk Minimization Measures for Blood Screening HIV-1 Nucleic Acid Amplification Technique Assays in Germany

2014 ◽  
Vol 41 (1) ◽  
pp. 8-8 ◽  
Author(s):  
Michael Chudy ◽  
Julia Kress ◽  
Jochen Halbauer ◽  
Margarethe Heiden ◽  
Markus B. Funk ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acid Amplification ◽  
Risk Minimization ◽  
Amplification Technique ◽  
Hiv 1 ◽  
Nucleic Acid Amplification Technique ◽  
Blood Screening

Quantitative nucleic acid amplification by digital PCR for clinical viral diagnostics

2016 ◽  
Vol 0 (0) ◽  
Author(s):  
Kuo Zhang ◽  
Guigao Lin ◽  
Jinming Li
Keyword(s):  
Nucleic Acid ◽  
Digital Pcr ◽  
Nucleic Acid Amplification ◽  
Target Sequence ◽  
The Past ◽  
Amplification Technique ◽  
Diagnostic Applications ◽  
Further Development ◽  
Nucleic Acid Amplification Technique ◽  
Viral Diagnostics

AbstractIn the past few years, interest in the development of digital PCR (dPCR) as a direct nucleic acid amplification technique for clinical viral diagnostics has grown. The main advantages of dPCR over qPCR include: quantification of nucleic acid concentrations without a calibration curve, comparable sensitivity, superior quantitative precision, greater resistance to perturbations by inhibitors, and increased robustness to the variability of the target sequence. In this review, we address the application of dPCR to viral nucleic acid quantification in clinical applications and for nucleic acid quantification standardization. Further development is required to overcome the current limitations of dPCR in order to realize its widespread use for viral load measurements in clinical diagnostic applications.

scholarly journals SIMPLIFIED DIAGNOSIS OF MALARIA INFECTION: GFM/PCR/ELISA A SIMPLIFIED NUCLEIC ACID AMPLIFICATION TECHNIQUE BY PCR/ELISA

1998 ◽  
Vol 40 (5) ◽  
pp. 333-334 ◽  
Author(s):  
Ricardo Luiz Dantas MACHADO ◽  
Denise Oliveira GARRET ◽  
Ipemida Sullayman ADAGU ◽  
David Charles WARHURST ◽  
Marinete Marins PÓVOA
Keyword(s):  
Nucleic Acid ◽  
Genetic Analysis ◽  
Blood Sample ◽  
Malaria Infection ◽  
Nucleic Acid Amplification ◽  
Sample Collection ◽  
Amplification Technique ◽  
The Cost ◽  
Nucleic Acid Amplification Technique

We report an adaptation of a technique for the blood sample collection (GFM) as well as for the extraction and amplification of Plasmodium DNA for the diagnosis of malaria infection by the PCR/ELISA. The method of blood sample collection requires less expertise and saves both time and money, thus reducing the cost by more than half. The material is also suitable for genetic analysis in either fresh or stored specimens prepared by this method.

scholarly journals World Health Organization International Standard To Harmonize Assays for Detection of Mycoplasma DNA

2015 ◽  
Vol 81 (17) ◽  
pp. 5694-5702 ◽  
Author(s):  
C. Micha Nübling ◽  
Sally A. Baylis ◽  
Kay-Martin Hanschmann ◽  
Thomas Montag-Lessing ◽  
Michael Chudy ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
World Health Organization ◽  
Nucleic Acid Amplification ◽  
World Health ◽  
Expert Committee ◽  
Content Type ◽  
International Standard ◽  
Amplification Technique ◽  
Health Organization ◽  
Nucleic Acid Amplification Technique

ABSTRACTNucleic acid amplification technique (NAT)-based assays (referred to here as NAT assays) are increasingly used as an alternative to culture-based approaches for the detection of mycoplasma contamination of cell cultures. Assay features, like the limit of detection or quantification, vary widely between different mycoplasma NAT assays. Biological reference materials may be useful for harmonization of mycoplasma NAT assays. An international feasibility study included lyophilized preparations of four distantly related mycoplasma species (Acholeplasma laidlawii,Mycoplasma fermentans,M. orale,M. pneumoniae) at different concentrations which were analyzed by 21 laboratories using 26 NAT assays with a qualitative, semiquantitative, or quantitative design. AnM. fermentanspreparation was shown to decrease the interassay variation when used as a common reference material. The preparation was remanufactured and characterized in a comparability study, and its potency (in NAT-detectable units) across different NATs was determined. The World Health Organization (WHO) Expert Committee on Biological Standardization (ECBS) established this preparation to be the “1st World Health Organization international standard for mycoplasma DNA for nucleic acid amplification technique-based assays designed for generic mycoplasma detection” (WHO Tech Rep Ser 987:42, 2014) with a potency of 200,000 IU/ml. This WHO international standard is now available as a reference preparation for characterization of NAT assays, e.g., for determination of analytic sensitivity, for calibration of quantitative assays in a common unitage, and for defining regulatory requirements in the field of mycoplasma testing.

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