A diagnostic test that uses isothermal amplification and lateral flow detection sdaA can detect tuberculosis in 60 min

2020 ◽  
Author(s):  
X. Wu ◽  
Y. Wang ◽  
Q. Yin ◽  
W. Jiao ◽  
L. Sun ◽  
...  
Keyword(s):  
Diagnostic Test ◽  
Lateral Flow ◽  
Isothermal Amplification ◽  
Flow Detection

scholarly journals Paper-Based RNA Extraction, in Situ Isothermal Amplification, and Lateral Flow Detection for Low-Cost, Rapid Diagnosis of Influenza A (H1N1) from Clinical Specimens

2015 ◽  
Vol 87 (15) ◽  
pp. 7872-7879 ◽  
Author(s):  
Natalia M. Rodriguez ◽  
Jacqueline C. Linnes ◽  
Andy Fan ◽  
Courtney K. Ellenson ◽  
Nira R. Pollock ◽  
...  
Keyword(s):  
Influenza A ◽  
Low Cost ◽  
Rna Extraction ◽  
Rapid Diagnosis ◽  
Lateral Flow ◽  
Isothermal Amplification ◽  
Clinical Specimens ◽  
Influenza A H1n1 ◽  
Flow Detection

scholarly journals Advanced Multiplex Loop Mediated Isothermal Amplification (mLAMP) Combined with Lateral Flow Detection (LFD) for Rapid Detection of Two Prevalent Malaria Species in India and Melting Curve Analysis

Diagnostics ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 32
Author(s):  
Supriya Sharma ◽  
Sandeep Kumar ◽  
Md.Zohaib Ahmed ◽  
Nitin Bhardwaj ◽  
Jaskirat Singh ◽  
...  
Keyword(s):  
Point Of Care ◽  
Melting Curve ◽  
Lateral Flow ◽  
Curve Analysis ◽  
Isothermal Amplification ◽  
Clinical Samples ◽  
Melting Curve Analysis ◽  
Loop Mediated Isothermal Amplification ◽  
Malaria Species ◽  
Flow Detection

Isothermal techniques with lateral flow detection have emerged as a point of care (POC) technique for malaria, a major parasitic disease in tropical countries such as India. Plasmodium falciparum and Plasmodium vivax are the two most prevalent malaria species found in the country. An advanced multiplex loop-mediated isothermal amplification (mLAMP) combined with a lateral flow dipstick (LFD) technique was developed for the swift and accurate detection of P. falciparum and P. vivax, overcoming the challenges of the existing RDTs (rapid diagnostic tests). A single set of LAMP primers with a biotinylated backward inner primer (BIP primer) was used for DNA amplification of both malaria species in a single tube. The amplified DNA was hybridized with fluorescein isothiocyanate (FITC) and digoxigenin-labelled DNA probes, having a complemented sequence for the P. falciparum and P. vivax genomes, respectively. A colour band appeared on two separate LFDs for P. falciparum and P. vivax upon running the hybridized solution over them. In total, 39 clinical samples were collected from ICMR-NIMR, New Delhi. Melting curve analysis, with cross primers for both species, was used to ascertain specificity, and the sensitivity was equated with a polymerase chain reaction (PCR). The results were visualized on the LFD for both species within 60 min. We found 100% sensitivity and specificity, when compared with a traditional PCR. Melting curve analysis of mLAMP revealed the lowest detection limit of 0.15 pg/μL from sample genomic DNA. The mLAMP-LFD assays could be a potential point of care (POC) tool for early diagnosis in non-laboratory conditions, with the convenience of a reduced assay time and the simple interpretation of results.

scholarly journals Isothermal Amplification and Lateral-Flow Assay for Detecting Crown-Gall-Causing Agrobacterium spp.

2017 ◽  
Vol 107 (9) ◽  
pp. 1062-1068 ◽  
Author(s):  
Skylar L. Fuller ◽  
Elizabeth A. Savory ◽  
Alexandra J. Weisberg ◽  
Jessica Z. Buser ◽  
Michael I. Gordon ◽  
...  
Keyword(s):  
Crown Gall ◽  
Molecular Detection ◽  
Lateral Flow ◽  
Isothermal Amplification ◽  
Molecular Diagnostic ◽  
Gram Negative Bacteria ◽  
Economic Costs ◽  
Crown Gall Disease ◽  
Oligonucleotide Primers ◽  
Flow Detection

Agrobacterium is a genus of soilborne gram-negative bacteria. Members carrying oncogenic plasmids can cause crown gall disease, which has significant economic costs, especially for the orchard and nursery industries. Early and rapid detection of pathogenic Agrobacterium spp. is key to the management of crown gall disease. To this end, we designed oligonucleotide primers and probes to target virD2 for use in a molecular diagnostic tool that relies on isothermal amplification and lateral-flow-based detection. The oligonucleotide tools were tested in the assay and evaluated for detection limit and specificity in detecting alleles of virD2. One set of primers that successfully amplified virD2 when used with an isothermal recombinase was selected. Both tested probes had detection limits in picogram amounts of DNA. Probe 1 could detect all tested pathogenic isolates that represented most of the diversity of virD2. Finally, the coupling of lateral-flow detection to the use of these oligonucleotide primers in isothermal amplification helped to reduce the onerousness of the process, and alleviated reliance on specialized tools necessary for molecular diagnostics. The assay is an advancement for the rapid molecular detection of pathogenic Agrobacterium spp.

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