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DNA dipstick

Ledicia Taboada, Ana Sánchez, Ricardo I Pérez-Martín, Carmen G Sotelo
Species-specific lateral flow dipstick (LFD) assays for the identification of Atlantic cod (Gadus morhua), Pacific cod (Gadus macrocephalus), Alaska pollock (Gadus chalcogrammus) and ling (Molva molva) in food products were developed. The method comprises a PCR system with four sets of specific primers, for each target species. This step was also devised to dual-labeling of PCR products with biotin and 6-FAM, which are then easily read on a lateral flow dipstick, upon which these products are immobilized by a fixed biotin-ligand and visualized with anti-FAM antibody-coated gold nanoparticles...
October 15, 2017: Food Chemistry
Rathina Kumar Shanmugakani, Yukihiro Akeda, Norihisa Yamamoto, Noriko Sakamoto, Hideharu Hagiya, Hisao Yoshida, Dan Takeuchi, Yo Sugawara, Takuya Kodera, Mitsuo Kawase, Warawut Laolerd, Narong Chaihongsa, Pitak Santanirand, Yoshikazu Ishii, Shigeyuki Hamada, Kazunori Tomono
A PCR-dipstick chromatography technique was designed and evaluated for differential identification of blaNDM, blaKPC, blaIMP, and blaOXA-48 carbapenemase genes directly in stool specimens within 2 h. It is a DNA-DNA hybridization-based detection system where PCR products can be easily interpreted by visual observation without electrophoresis. The PCR-dipstick showed high sensitivity (93.3%) and specificity (99.1%) in directly detecting carbapenemase genes in stool specimens compared with multiplex PCR for genomic DNA of the isolates from those stool specimens...
June 2017: Antimicrobial Agents and Chemotherapy
Phiaw Chong Foo, Yean Yean Chan, Maizan Mohamed, Weng Kin Wong, A B Nurul Najian, Boon Huat Lim
This study highlighted the development of a four target nitrocellulose-based nucleic acid lateral flow immunoassay biosensor in a dry-reagent strip format for interpretation of double-labelled double-stranded amplicons from thermostabilised triplex loop-mediated isothermal amplification assay. The DNA biosensor contained two test lines which captured biotin and texas red labelled amplicons; a LAMP internal amplification control line that captured digoxigenin labelled amplicon; and a chromatography control line that validated the functionality of the conjugated gold nanoparticles and membrane...
May 8, 2017: Analytica Chimica Acta
Zorica Dakić, Henrik Vedel Nielsen, Milorad Pavlović, Jasmina Poluga, Goran Stevanović, Lidija Lavadinović, Branko Milošević, Mijomir Pelemiš, Aleksandar Urošević, Snežana Jovanović, Christen Rune Stensvold
INTRODUCTION: Retrospective molecular identification of Leishmania parasites in two patients with visceral leishmaniasis (VL) previously treated in Serbia was carried out. DNA was isolated from unstained bone marrow smears (BMSs) kept for 11 and 8 years. Genus-specific real-time PCR was combined with conventional PCR and sequencing for detection and species identification. CASE PRESENTATION: In 2003, a 40-year-old Serbian male was admitted to the Clinical Centre of Serbia (CCS) with fever, sweating, fatigue and splenomegaly, which developed over a period of 7 weeks...
October 2016: JMM Case Reports
Ratchanida Kumvongpin, Patcharee Jearanaikoon, Chotchana Wilailuckana, Nattaya Sae-Ung, Prinya Prasongdee, Sakda Daduang, Metee Wongsena, Patcharee Boonsiri, Wansika Kiatpathomchai, Sukumarn Sanersak Swangvaree, Alisa Sandee, Jureerut Daduang
Cervical cancer is the third highest cause of death in developing countries and most commonly results from high‑risk human papillomavirus (HR‑HPV) infection. Among HR‑HPV genotypes, HPV16 and HPV18 are the most prevalent in cervical cancers. Therefore, the present study aimed to develop a detection assay for HPV16 and HPV18 infection using loop‑mediated isothermal amplification (LAMP) with lateral flow dipstick (LFD) tests. This assay is a simplified, user‑friendly method for the visual detection of HPV genotypes...
May 2017: Molecular Medicine Reports
Gad Baneth, Daniel Yasur-Landau, Matan Gilad, Yaarit Nachum-Biala
BACKGROUND: Infection and clinical disease associated with Leishmania major and Leishmania tropica, two common agents of human cutaneous leishmaniosis, have rarely been reported in dogs. This study describes dogs infected with these Leishmania spp. prevalent in the Middle East and North Africa, and compares the serological response of dogs infected with Leishmania infantum, L. major or L. tropica to whole promastigote antigen enzyme-linked immunosorbent assay (ELISA) of each species and to rK39 dipstick...
March 13, 2017: Parasites & Vectors
Debapriya Mazumdar, Tian Lan, Yi Lu
Real-time, on-site detection and quantification of different trace analytes is a challenge that requires both searching a general class of molecules to recognize a broad range of contaminants and translating this recognition to easily detectable signals. Functional nucleic acids, which include DNAzymes (DNA with catalytic activity) and aptamers (nucleic acids that bind an analyte), are ideal candidates for the target recognition. These nucleic acids can be selected by a combinatorial biology method called in vitro selection to interact with a particular analyte with high specificity and sensitivity...
2017: Methods in Molecular Biology
Michelle C Stanton, Moses Adriko, Moses Arinaitwe, Alison Howell, Juliet Davies, Gillian Allison, E James LaCourse, Edridah Muheki, Narcis B Kabatereine, J Russell Stothard
BACKGROUND: Intestinal schistosomiasis is of public health importance in Uganda but communities living above 1400 m are not targeted for control as natural transmission is thought unlikely. To assess altitudinal boundaries and at-risk populations, conjoint malacological and epidemiological surveys were undertaken on Mount Elgon (1139 m-3937 m), in Fort Portal crater lakes and in the Rwenzori Mountains (1123 m-4050 m). METHODS: Seventy freshwater habitats [Mount Elgon (37), Fort Portal crater lakes (23), Rwenzori Mountains (8) and Lake Albert (2)] were inspected for Biomphalaria species...
February 6, 2017: Infectious Diseases of Poverty
Po-An Tu, Jia-Shian Shiu, Shu-Hwae Lee, Victor Fei Pang, De-Chi Wang, Pei-Hwa Wang
Caprine arthritis-encephalitis (CAE) in goats is a complex disease syndrome caused by a lentivirus. This persistent viral infection results in arthritis in adult goats and encephalitis in lambs. The prognosis for the encephalitic form is normally poor, and this form of the disease has caused substantial economic losses for goat farmers. Hence, a more efficient detection platform based on recombinase polymerase amplification (RPA) and a lateral flow dipstick (LFD) was developed in the present study for detecting the proviral DNA of caprine arthritis-encephalitis virus (CAEV)...
May 2017: Journal of Virological Methods
Hai-Long Huang, Peng Zhu, Cheng-Xu Zhou, Shan He, Xiao-Jun Yan
The aim of this study was to develop a loop-mediated isothermal amplification (LAMP) combined with a chromatographic lateral flow dipstick (LFD) assay to rapidly and specifically detect the Karlodinium veneficum ITS gene. Four groups of LAMP primers were specially designed to target the K. veneficum ITS gene. The LAMP-LFD detection limit was 7.4pg/μL (approximately 6.5cells/mL) of K. veneficum genomic DNA and was 10 times more sensitive than standard PCR. The LAMP-LFD method exhibited high specificity and accurately identified K...
February 2017: Harmful Algae
Kristofer J Haushalter, Srinivas Vetcha, Robert C Haushalter
Lateral flow or dipstick assays (e.g., home pregnancy tests), where an analyte solution is drawn through a porous membrane and is detected by localization onto a capture probe residing at a specific site on the flow strip, are the most commonly and extensively used type of diagnostic assay. However, after over 30 years of use, these assays are constrained to measuring one or a few analytes at a time. Here, we describe a completely general method, in which any single-plex lateral flow assay is transformed into a multiplex assay capable of measuring an arbitrarily large number of analytes simultaneously...
October 31, 2016: ACS Omega
Suganya Yongkiettrakul, Jantana Kampeera, Wanwisa Chareanchim, Roonglawan Rattanajak, Wichai Pornthanakasem, Wansika Kiatpathomchai, Darin Kongkasuriyachai
The significant strides made in reducing global malaria burden over the past decades are being threatened by the emergence of multi-drug resistant malaria. Mechanisms of resistance to several classes of antimalarial drugs have been linked to key mutations in the Plasmodium falciparum genes. Pyrimethamine targets the dihydrofolate reductase of the bifunctional dihydrofolate reductase thymidylate synthase (DHFR-TS), and specific point mutations in the dhfr-ts gene have been assigned to resistant phenotypes. Several molecular methods are available to detect the mutant genotypes including DNA sequencing and PCR-based methods...
February 2017: Parasitology International
Shankar Mandal, Sangeetha Selvam, Prakash Shrestha, Hanbin Mao
Sensitivity of biosensors is set by the dissociation constant (Kd) between analytes and probes. Although potent amplification steps can be accommodated between analyte recognition and signal transduction in a sensor to improve the sensitivity 4-6 orders of magnitude below Kd, they compromise temporal resolution. Here, we demonstrated mechanochemical sensing that broke the Kd limit by 9 orders of magnitude for Hg detection without amplifications. Analogous to trawl fishing, we introduced multiple Hg binding units (thymine (T)-T pairs) in a molecular trawl made of two poly-T strands...
October 4, 2016: Analytical Chemistry
Kui Sun, Weiwei Xing, Xinling Yu, Wenliang Fu, Yuanyuan Wang, Minji Zou, Zhihong Luo, Donggang Xu
BACKGROUND: With the continuous decline in prevalence and intensity of Schistosoma japonicum infection in China, more accurate and sensitive methods suitable for field detection become much needed for schistosomiasis control. Here, a novel rapid and visual detection method based on the combination of recombinase polymerase amplification (RPA) and lateral flow dipstick (LFD) was developed to detect S. japonicum DNA in fecal samples. RESULTS: The LFD-RPA assay targeting SjR2 could detect 5 fg S...
2016: Parasites & Vectors
Kawin Nawattanapaiboon, Photchanathorn Prombun, Pitak Santanirand, Apirom Vongsakulyanon, Toemsak Srikhirin, Boonsong Sutapun, Wansika Kiatpathomchai
This study reports loop-mediated isothermal amplification (LAMP) for rapid detection of methicillin-resistant Staphylococcus aureus from direct clinical specimens. Four primers including outer and inner primers were specifically designed on the two target sequences-femB to identify S. aureus and mecA to identify antibiotic-resistant gene. Reference strains including various species of gram-positive/gram-negative isolates were used to evaluate and optimize LAMP assays. The optimum LAMP condition was found at 63°C within 70 min assay time (include hybridization with FITC probe for 5 min and further 5 min for reading the results on the lateral flow dipstick)...
September 2016: Journal of Clinical Laboratory Analysis
Yimin Yang, Qun Li, Suhua Wang, Xueqiu Chen, Aifang Du
Babesia spp. are apicomplexan protozoan parasites of the red blood cells of mammals and are transmitted by ticks. Bovine babesiosis mainly caused by Babesia bovis and Babesia bigemina occurs worldwide, which is a great threat to animal health. Microscopy examination is a gold standard for the diagnosis of babesiosis. However, its sensitivity is too low. This study was conducted to establish a simple, efficient and fast LAMP-LFP method used for early diagnosis of animal babesiosis. LAMP was developed with a set of four primers targeting and amplifying six distinct regions of cytochrome b gene of Babesia spp...
March 30, 2016: Veterinary Parasitology
Lingyang Tian, Takuichi Sato, Kousuke Niwa, Mitsuo Kawase, Gen Mayanagi, Jumpei Washio, Nobuhiro Takahashi
The onset of plaque-mediated disease, including dental caries and periodontal diseases, is highly associated with compositional change of the resident microflora from the ecological perspective. As specific bacterial profiles have been linked to different disease stages, microbial compositional measurements might therefore have great value for clinical diagnosis. Previously we have reported a dry-reagent strip biosensor-PCR-dipstick DNA chromatography, which utilized molecular recognition of oligonucleotides and biotin-streptavidin, and the optical property of colored microspheres, for semiquantifying a five-membered subgroup of caries-associated bacterial species in supragingival plaque from healthy coronal surfaces of teeth...
2016: Biomedical Research
Umi Rubiah Sastu, Noor Rain Abdullah, Nor Azrina Norahmad, Muhammad Nor Farhan Saat, Prem Kumar Muniandy, Jenarun Jelip, Moizin Tikuson, Norsalleh Yusof, Hasidah Mohd Sidek
BACKGROUND: Malaria cases persist in some remote areas in Sabah and Sarawak despite the ongoing and largely successful malaria control programme conducted by the Vector Borne Disease Control Programme, Ministry Of Health, Malaysia. Point mutations in the genes that encode the two enzymes involved in the folate biosynthesis pathway, dihydrofolate reductase (DHFR) and dihydropteroate synthase (DHPS) enzymes confer resistance to pyrimethamine and sulfadoxine respectively, in both Plasmodium falciparum and P...
February 5, 2016: Malaria Journal
Nikolaos Fountoglou, Margarita Petropoulou, Alexandra Iliadi, Theodore K Christopoulos, Penelope C Ioannou
Thrombosis is considered as the most typical example of multigenic/multifactorial disorder. The three most common genetic risk factors for thrombotic disorders are the G1691A mutation in factor V gene (FV Leiden), the G20210Α mutation in prothrombin gene (FII), and the C677T mutation in the methylenetetrahydrofolate reductase (MTHFR) gene. An additional panel of biomarkers predisposing for thrombotic events includes the H1299R variant in factor V gene (HR2), A1298C variant in MTHFR gene, the V34L mutation in fibrinogen stabilizing factor XIII (FXIII) gene as well as the 4G/5G polymorphism in plasminogen activator inhibitor type-1 (PAI-1) gene...
March 2016: Analytical and Bioanalytical Chemistry
A B Nurul Najian, E A R Engku Nur Syafirah, Nabilah Ismail, Maizan Mohamed, Chan Yean Yean
In recent years extensive numbers of molecular diagnostic methods have been developed to meet the need of point-of-care devices. Efforts have been made towards producing rapid, simple and inexpensive DNA tests, especially in the diagnostics field. We report on the development of a label-based lateral flow dipstick for the rapid and simple detection of multiplex loop-mediated isothermal amplification (m-LAMP) amplicons. A label-based m-LAMP lateral flow dipstick assay was developed for the simultaneous detection of target DNA template and a LAMP internal control...
January 15, 2016: Analytica Chimica Acta
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