scholarly journals Nucleic Acid-Based Lateral Flow Biosensor for Salmonella Typhi and Salmonella Paratyphi: A Detection in Stool Samples of Suspected Carriers

Diagnostics ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 700
Author(s):  
Zulkiply Nor Amalina ◽  
Muhammad Fazli Khalid ◽  
Sjafri Faizul Rahman ◽  
Muhamad Nuramin Ahmad ◽  
Mohamad Ahmad Najib ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Rapid Detection ◽  
Detection System ◽  
Flow Analysis ◽  
Fluorescein Isothiocyanate ◽  
Salmonella Typhi ◽  
Lateral Flow ◽  
Surveillance Program ◽  
Biotin Binding ◽  
Stool Samples

A multiplex rapid detection system, based on a PCR-lateral flow biosensor (mPCR-LFB) was developed to identify Salmonella Typhi and Salmonella Paratyphi A from suspected carriers. The lower detection limit for S. Typhi and S. Paratyphi A was 0.16 and 0.08 ng DNA equivalent to 10 and 102 CFU/mL, respectively. Lateral flow biosensor was used for visual detection of mPCR amplicons (stgA, SPAint, ompC, internal amplification control) by labeling forward primers with fluorescein-isothiocyanate (FITC), Texas Red, dinitrophenol (DNP) and digoxigenin (DIG) and reverse primers with biotin. Binding of streptavidin-colloidal gold conjugate with the amplicons resulted in formation of a red color dots on the strip after 15–20 min of sample exposure. The nucleic acid lateral flow analysis of the mPCR-LFB was better in sensitivity and more rapid than the conventional agarose gel electrophoresis. Moreover, the mPCR-LFB showed 100% sensitivity and specificity when evaluated with stools spiked with 100 isolates of Salmonella genus and other bacteria. A prospective cohort study on stool samples of 1176 food handlers in outbreak areas (suspected carriers) resulted in 23 (2%) positive for S. Typhi. The developed assay has potential to be used for rapid detection of typhoid carriers in surveillance program.

scholarly journals Quantum dots nanoparticle-based lateral flow assay for rapid detection of Mycobacterium species using anti-FprA antibodies

2012 ◽  
Vol 2 (1) ◽  
pp. 5 ◽  
Author(s):  
Fabio Cimaglia ◽  
Alessandro Aliverti ◽  
Maurizio Chiesa ◽  
Palmiro Poltronieri ◽  
Enrico De Lorenzis ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Quantum Dots ◽  
Monoclonal Antibodies ◽  
Rapid Detection ◽  
Detection System ◽  
Lateral Flow ◽  
Fluorescence Signal ◽  
Human Pathogens ◽  
Detection Zone ◽  
Diagnostic Investigation

A lateral flow (LF) device combined with quantum dots (QDs) technology was developed for rapid detection of a specific mycobacterial flavoprotein reductase (<em>fprA</em>). In order to develop the LF assay based on a double-antibody sandwich format, two monoclonal antibodies recognizing different epitopes located in separated <em>fprA</em> domains were identified. The first monoclonal antibody was immobilized onto the detection zone of a porous nitrocellulose membrane, whereas another monoclonal antibody was conjugated to QDs nanoparticles as a detection system. Using these monoclonal antibodies we recorded a good fluorescence signal, the intensity of which was directly proportional to the concentration of <em>fprA</em> protein. The use of antibodies conjugated with fluorescent semiconductor QDs via biotin-streptavidin bridge, allowed the detection of <em>fprA</em> protein at concentrations as low as 12.5 pg/μL in less than 10 min. The reported technology could be useful in the diagnostic investigation of <em>Mycobacterium tuberculosis</em> and other human pathogens in clinical specimens.

Development of a nucleic acid lateral flow immunoassay (NALFIA) for reliable, simple and rapid detection of the methicillin resistance genes mecA and mecC

2017 ◽  
Vol 200 ◽  
pp. 101-106 ◽  
Author(s):  
Constanze Seidel ◽  
Sonja Peters ◽  
Erik Eschbach ◽  
Andrea T. Feßler ◽  
Boris Oberheitmann ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Resistance Genes ◽  
Rapid Detection ◽  
Methicillin Resistance ◽  
Lateral Flow ◽  
Lateral Flow Immunoassay

scholarly journals Highly Specific and Sensitive Detection of Yersinia pestis by Portable Cas12a-UPTLFA Platform

2021 ◽  
Vol 12 ◽  
Author(s):  
Yang You ◽  
Pingping Zhang ◽  
Gengshan Wu ◽  
Yafang Tan ◽  
Yong Zhao ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Yersinia Pestis ◽  
Rapid Detection ◽  
Detection Method ◽  
Detection System ◽  
Point Of Care ◽  
High Sensitivity ◽  
Nucleic Acid Detection ◽  
Great Promise ◽  
High Background

The recent discovery of collateral cleavage activity of class-II clustered regularly interspaced short palindromic repeats–CRISPR-associated protein (CRISPR-Cas) makes CRISPR-based diagnosis a potential high-accuracy nucleic acid detection method. Colloidal gold-based lateral flow immunochromatographic assay (LFA), which has been combined with CRISPR/Cas-based nucleic detection, usually associates with drawbacks of relative high background and the subjectivity in naked-eye read-out of the results. Here, we developed a novel system composed of Cas12a-based nucleic acid detection and up-converting phosphor technology (UPT)-based LFA (UPT–LFA), termed Cas12a-UPTLFA. We further demonstrated the utility of this platform in highly sensitive and specific detection of Yersinia pestis, the causative agent of the deadly plague. Due to high infectivity and mortality, as well as the potential to be misused as bioterrorism agent, a culture-free, ultrasensitive, specific, and rapid detection method for Y. pestis has long been desired. By incorporating isothermal recombinase polymerase amplification, the Cas12a-UPTLFA we established can successfully detect genomic DNA of Y. pestis as low as 3 attomolar (aM) and exhibited high sensitivity (93.75%) and specificity (90.63%) for detection of spiked blood samples with a detection limit of 102 colony-forming unit per 100 μl of mouse blood. With a portable biosensor, Cas12a-UPTLFA assay can be operated easily by non-professional personnel. Taken together, we have developed a novel Cas12a-UPTLFA platform for rapid detection of Y. pestis with high sensitivity and specificity, which is portable, not expensive, and easy to operate as a point-of-care method. This detection system can easily be extended to detect other pathogens and holds great promise for on-site detection of emerging infectious pathogens.

scholarly journals Rapid Detection of Pathogens in Wound Exudate via Nucleic Acid Lateral Flow Immunoassay

Biosensors ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 74
Author(s):  
Anna Brunauer ◽  
René D. Verboket ◽  
Daniel M. Kainz ◽  
Felix von Stetten ◽  
Susanna M. Früh
Keyword(s):  
Nucleic Acid ◽  
Rapid Detection ◽  
Direct Detection ◽  
Point Of Care ◽  
False Negative ◽  
Opportunistic Pathogen ◽  
Lateral Flow ◽  
Lateral Flow Immunoassay ◽  
Wound Fluid ◽  
Wound Exudate

The rapid detection of pathogens in infected wounds can significantly improve the clinical outcome. Wound exudate, which can be collected in a non-invasive way, offers an attractive sample material for the detection of pathogens at the point-of-care (POC). Here, we report the development of a nucleic acid lateral flow immunoassay for direct detection of isothermally amplified DNA combined with fast sample preparation. The streamlined protocol was evaluated using human wound exudate spiked with the opportunistic pathogen Pseudomonas aeruginosa that cause severe health issues upon wound colonization. A detection limit of 2.1 × 105 CFU per mL of wound fluid was achieved, and no cross-reaction with other pathogens was observed. Furthermore, we integrated an internal amplification control that excludes false negative results and, in combination with the flow control, ensures the validity of the test result. The paper-based approach with only three simple hands-on steps has a turn-around time of less than 30 min and covers the complete analytical process chain from sample to answer. This newly developed workflow for wound fluid diagnostics has tremendous potential for reliable pathogen POC testing and subsequent target-oriented therapy.

Rapid detection of Rotavirus antigen in stool sample of acute diarrheic children

2012 ◽  
Vol 6 (1) ◽  
pp. 11-13
Author(s):  
Sushmita Roy ◽  
S.M. Shamsuzzaman ◽  
K.Z. Mamun
Keyword(s):  
Stool Sample ◽  
Rapid Detection ◽  
Cross Sectional Study ◽  
Rapid Diagnosis ◽  
Age Group ◽  
College Hospital ◽  
Cross Sectional ◽  
Rotavirus Infection ◽  
Medical College ◽  
Stool Samples

Rotavirus is one of the leading causes of pediatric diarrhea globally. Accurate and rapid diagnosis of Rotavirus diarrhea should reduce unnecessary use of antibiotics and ultimately reduce drug resistance. Study was designed for rapid diagnosis of Rotavirus antigen in stool sample by ICT (Immunochromatographic test) as well as to observe the seasonal variation of rotavirus infection. This cross sectional study was carried out in the department of Microbiology, Dhaka Medical College from January 2011 to December 2011. Eighty stool samples were collected from Dhaka Shishu Hospital and Dhaka Medical College Hospital. All samples were tested for rotavirus antigen by ICT. Among 80 patients, 42 (52.5%) samples were positive for rotavirus antigen. Among these 42 positive samples, 30 (71.43%) were from 0-12 months of age group, 10 (23.81%) from 13 to 24 months of age group and rest 2 (4.76%) from 25 to 36 months of age group. Rotavirus Ag was detected in stool samples from January to April and another peak episode from October to December. Considering the importance of Rotavirus associated diarrhea, rapid detection of Rotavirus infection in human is substantially needed and should be routinely practiced.DOI: http://dx.doi.org/10.3329/bjmm.v6i1.19354 Bangladesh J Med Microbiol 2012; 06(01): 11-13

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