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https://www.readbyqxmd.com/read/28098507/development-and-evaluation-of-a-loop-mediated-isothermal-amplification-lamp-assay-for-the-detection-of-haemonchus-contortus-in-goat-fecal-samples
#1
Xin Yang, Mingwei Qi, Zongze Zhang, Chong Gao, Chunqun Wang, Weiqiang Lei, Li Tan, Junlong Zhao, Rui Fang, Min Hu
Haemonchus contortus is one of the most significant strongylid nematodes infecting small ruminants and causes great economic losses to the livestock industry worldwide. Accurate diagnosis of H. contortus is crucial to control the infection. Traditional microscopic examinations are the most common methods for the diagnosis of H. contortus, but they are time-consuming and inaccurate. Molecular methods based on PCR are more accurate, but need expensive machines usually only used in the laboratory. Loop-mediated isothermal amplification (LAMP) is a rapid, simple, specific and sensitive method that has been widely used to detect viruses, bacteria and parasites...
January 18, 2017: Journal of Parasitology
https://www.readbyqxmd.com/read/28096624/genetic-variability-and-discrimination-of-low-doses-of-toxocara-spp-from-public-areas-soil-inferred-by-loop-mediated-isothermal-amplification-assay-as-a-field-friendly-molecular-tool
#2
Maryam Ozlati, Adel Spotin, Abbas Shahbazi, Mahmoud Mahami-Oskouei, Teimour Hazratian, Mohammad Adibpor, Ehsan Ahmadpour, Afsaneh Dolatkhah, Paria Khoshakhlagh
ABSTRACT: Aim: One of the main diagnostic problems of conventional polymerase chain reaction (PCR) is indiscrimination of low parasitic loads in soil samples. The aim of this study is to determine the genetic diversity and identification of Toxocara spp. from public areas soil inferred by loop-mediated isothermal amplification (LAMP) assay. MATERIALS AND METHODS: A total of 180 soil samples were collected from various streets and public parks of northwest Iran. The DNA of recovered Toxocara eggs were extracted and amplified by PCR and LAMP following ZnSO4 flotation technique...
December 2016: Veterinary World
https://www.readbyqxmd.com/read/28096435/regulation-of-aldosterone-secretion-by-mineralocorticoid-receptor-mediated-signaling
#3
Cherish Chong, Anis Hamid, Tham Yao, Amanda Garza, Luminita Pojoga, Gail K Adler, Jose R Romero, Gordon H Williams
We posit the existence of a paracrine/autocrine negative feedback loop, mediated by the mineralocorticoid receptor (MR), regulating aldosterone secretion. To assess this hypothesis we asked whether altering MR activity in zona glomerulosa (ZG) cells affects aldosterone production. To this end, we studied ex vivo ZG cells isolated from male Wistar rats fed chow containing either high (1.6% Na+ [HS]) or low (0.03% Na+ [LS]) amount of sodium. Western blot analyses demonstrated that MR was present in both the ZG and zona fasciculata/zona reticularis (ZF/ZR/ZR)...
January 17, 2017: Journal of Endocrinology
https://www.readbyqxmd.com/read/28095790/comparison-of-loop-mediated-isothermal-amplification-assay-and-smear-microscopy-with-culture-for-the-diagnostic-accuracy-of-tuberculosis
#4
Baye Gelaw, Yitayal Shiferaw, Marta Alemayehu, Abate Assefa Bashaw
BACKGROUND: Tuberculosis (TB) caused by Mycobacterium tuberculosis is one of the leading causes of death from infectious diseases worldwide. Sputum smear microscopy remains the most widely available pulmonary TB diagnostic tool particularly in resource limited settings. A highly sensitive diagnostic with minimal infrastructure, cost and training is required. Hence, we assessed the diagnostic performance of Loop-mediated isothermal amplification (LAMP) assay in detecting M.tuberculosis infection in sputum sample compared to LED fluorescent smear microscopy and culture...
January 17, 2017: BMC Infectious Diseases
https://www.readbyqxmd.com/read/28095434/field-evaluation-of-a-high-throughput-loop-mediated-isothermal-amplification-test-for-the-detection-of-asymptomatic-plasmodium-infections-in-zanzibar
#5
Berit Aydin-Schmidt, Ulrika Morris, Xavier C Ding, Irina Jovel, Mwinyi I Msellem, Daniel Bergman, Atiqul Islam, Abdullah S Ali, Spencer Polley, Iveth J Gonzalez, Andreas Mårtensson, Anders Björkman
BACKGROUND: New field applicable diagnostic tools are needed for highly sensitive detection of residual malaria infections in pre-elimination settings. Field performance of a high throughput DNA extraction system for loop mediated isothermal amplification (HTP-LAMP) was therefore evaluated for detecting malaria parasites among asymptomatic individuals in Zanzibar. METHODS: HTP-LAMP performance was evaluated against real-time PCR on 3008 paired blood samples collected on filter papers in a community-based survey in 2015...
2017: PloS One
https://www.readbyqxmd.com/read/28093275/reverse-transcription-loop-mediated-isothermal-amplification-for-rapid-detection-of-respiratory-syncytial-virus-directly-from-nasopharyngeal-swabs
#6
Johannes Hoos, Rebecca Marie Peters, Julia Tabatabai, Jürgen Grulich-Henn, Paul Schnitzler, Johannes Pfeil
Respiratory syncytial virus (RSV) is a common cause of lower respiratory tract infection in young infants and a major cause of nosocomial infection in pediatric care. Currently available RSV point-of-care tests are of limited sensitivity and relatively expensive. We developed and evaluated a novel RSV rapid test for use at point-of-care, based on reverse-transcription loop-mediated isothermal amplification (RT-LAMP) for direct testing of nasopharyngeal swab specimens. RT-LAMP can detect RSV within 30min, without the need for RNA extraction...
January 13, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28092481/the-effect-of-main-urine-inhibitors-on-the-activity-of-different-dna-polymerases-in-loop-mediated-isothermal-amplification
#7
Jekaterina Jevtuševskaja, Katrin Krõlov, Indrek Tulp, Ülo Langel
BACKGROUND: The use of rapid amplification methods to detect pathogens in biological samples is mainly limited by the amount of pathogens present in the sample and the presence of inhibiting substances. Inhibitors can affect the amplification efficiency by either binding to the polymerase, interacting with the DNA, or interacting with the polymerase during primer extension. Amplification is performed using DNA polymerase enzymes and even small changes in their activity can influence the sensitivity and robustness of molecular assays Methods: The main purpose of this research was to examine which compounds present in urine inhibit polymerases with strand displacement activity...
January 16, 2017: Expert Review of Molecular Diagnostics
https://www.readbyqxmd.com/read/28092385/visual-detection-of-multiple-genetically-modified-organisms-in-a-capillary-array
#8
Ning Shao, Jianwei Chen, Jiaying Hu, Rong Li, Dabing Zhang, Shujuan Guo, Junhou Hui, Peng Liu, Litao Yang, Sheng-Ce Tao
There is an urgent need for rapid, low-cost multiplex methodologies for the monitoring of genetically modified organisms (GMOs). Here, we report a C[combining low line]apillary A[combining low line]rray-based L[combining low line]oop-mediated isothermal amplification for M[combining low line]ultiplex visual detection of nucleic acids (CALM) platform for the simple and rapid monitoring of GMOs. In CALM, loop-mediated isothermal amplification (LAMP) primer sets are pre-fixed to the inner surface of capillaries...
January 16, 2017: Lab on a Chip
https://www.readbyqxmd.com/read/28088238/lymph-node-pooling-a-feasible-and-efficient-method-of-lymph-node-molecular-staging-in-colorectal-carcinoma
#9
Natalia Rakislova, Carla Montironi, Iban Aldecoa, Eva Fernandez, Josep Antoni Bombi, Mireya Jimeno, Francesc Balaguer, Maria Pellise, Antoni Castells, Miriam Cuatrecasas
BACKGROUND: Pathologic lymph node staging is becoming a deficient method in the demanding molecular era. Nevertheless, the use of more sensitive molecular analysis for nodal staging is hampered by its high costs and extensive time requirements. Our aim is to take a step forward in colon cancer (CC) lymph node (LN) pathology diagnosis by proposing a feasible and efficient molecular method in routine practice using reverse transcription loop-mediated isothermal amplification (RT-LAMP). RESULTS: Molecular detection of tumor cytokeratin 19 (CK19) mRNA with RT-LAMP was performed in 3206 LNs from 188 CC patients using two methods: individual analysis of 1449 LNs from 102 patients (individual cohort), and pooled LN analysis of 1757 LNs from 86 patients (pooling cohort)...
January 14, 2017: Journal of Translational Medicine
https://www.readbyqxmd.com/read/28086789/detection-of-plasmodium-knowlesi-plasmodium-falciparum-and-plasmodium-vivax-using-loop-mediated-isothermal-amplification-lamp-in-a-co-endemic-area-in-malaysia
#10
Kim A Piera, Ammar Aziz, Timothy William, David Bell, Iveth J González, Bridget E Barber, Nicholas M Anstey, Matthew J Grigg
BACKGROUND: Plasmodium knowlesi is the most common cause of malaria in Malaysia. However, microscopic diagnosis is inaccurate and rapid diagnostic tests (RDTs) are insufficiently sensitive. PCR is sensitive and specific but not feasible at a district level. Loop-mediated isothermal amplification (LAMP) shows potential with only basic requirements. A commercially available LAMP assay, the Eiken Loopamp™ MALARIA Pan Detection kit, is sensitive for Plasmodium falciparum and Plasmodium vivax, but has not previously been evaluated for P...
January 13, 2017: Malaria Journal
https://www.readbyqxmd.com/read/28074711/a-comparison-of-the-reliability-of-two-gene-targets-in-loop-mediated-isothermal-amplification-assays-for-detecting-leptospiral-dna-in-canine-urine
#11
Fabio Gentilini, Renato Giulio Zanoni, Elisa Zambon, Maria Elena Turba
We compared 2 novel loop-mediated isothermal amplification (LAMP) assays that target either the 16S ribosomal RNA ( rrs) gene or the gene encoding a 32-kDa leptospiral lipoprotein ( lipL32) in order to assess the effect of the target on the accuracy of the LAMP assays. The most sensitive assay was the rrs assay with a limit of detection (LOD) of 1.2 × 10(1) genome equivalents per reaction. The novel lipL32 assay showed an LOD of 1.2 × 10(2) genome equivalents per reaction. Both assays showed adequate specificity when tested against a collection of bacteria commonly found in voided canine urine...
January 2017: Journal of Veterinary Diagnostic Investigation
https://www.readbyqxmd.com/read/28073898/two-stage-isothermal-enzymatic-amplification-for-concurrent-multiplex-molecular-detection
#12
Jinzhao Song, Changchun Liu, Michael G Mauk, Shelley C Rankin, James B Lok, Robert M Greenberg, Haim H Bau
BACKGROUND: The wide array of pathogens responsible for infectious diseases makes it difficult to identify causative pathogens with single-plex tests. Although multiplex PCR detects multiple targets, it is restricted to centralized laboratories, which delays test results or makes multiplexing unavailable, depriving healthcare providers of critical, real-time information. METHODS: To address the need for point-of-care (POC) highly multiplexed tests, we propose the 2-stage, nested-like, rapid (<40 min) isothermal amplification assay, dubbed rapid amplification (RAMP)...
January 10, 2017: Clinical Chemistry
https://www.readbyqxmd.com/read/28070163/development-of-reverse-transcription-loop-mediated-isothermal-amplification-for-simple-and-rapid-detection-of-promyelocytic-leukemia-retinoic-acid-receptor-%C3%AE-mrna
#13
Yuki Hashimoto, Yuki Hatayama, Nao Kojima, Shota Morishita, Satoko Matsumoto, Yuzuru Hosoda, Ayako Hara, Toru Motokura
BACKGROUND: Acute promyelocytic leukemia (APL) is a disease characterized by expression of Promyelocytic Leukemia-Retinoic Acid Receptor α (PML-RARα) chimeric mRNA. Although APL is curable, early death due to hemorrhage is a major problem. Here, we report the development of a simple and rapid diagnostic method for APL based on reverse transcription loop-mediated isothermal amplification (RT-LAMP). METHODS: An RT-LAMP primer set was designed to detect three types of PML-RARα mRNA in a single reaction...
December 2016: Yonago Acta Medica
https://www.readbyqxmd.com/read/28069610/an-incoherent-feed-forward-loop-mediates-robustness-and-tunability-in-a-plant-immune-network
#14
Akira Mine, Tatsuya Nobori, Maria C Salazar-Rondon, Thomas M Winkelmüller, Shajahan Anver, Dieter Becker, Kenichi Tsuda
Immune signaling networks must be tunable to alleviate fitness costs associated with immunity and, at the same time, robust against pathogen interferences. How these properties mechanistically emerge in plant immune signaling networks is poorly understood. Here, we discovered a molecular mechanism by which the model plant species Arabidopsis thaliana achieves robust and tunable immunity triggered by the microbe-associated molecular pattern, flg22. Salicylic acid (SA) is a major plant immune signal molecule...
January 9, 2017: EMBO Reports
https://www.readbyqxmd.com/read/28063000/evaluation-of-loop-mediated-isothermal-amplification-for-the-rapid-identification-of-bacteria-and-resistance-determinants-in-positive-blood-cultures
#15
J Rödel, J A Bohnert, S Stoll, L Wassill, B Edel, M Karrasch, B Löffler, W Pfister
The use of molecular assays to rapidly identify pathogens and resistance genes directly from positive blood cultures (BCs) contribute to shortening the time required for the diagnosis of bloodstream infections. In this work, loop-mediated isothermal amplification (LAMP) assays have been examined for their potential use in BC diagnosis. Three different assays were applied. The commercially available eazyplex® MRSA test detects Staphylococcus aureus, S. epidermidis, mecA, and mecC. Two in-house assays [Gram-positive (GP) and Gram-negative (GN)] have been developed for the detection of streptococci, enterococci, vanA, vanB, Pseudomonas spp...
January 6, 2017: European Journal of Clinical Microbiology & Infectious Diseases
https://www.readbyqxmd.com/read/28062857/positive-feedback-loop-mediated-by-protein-phosphatase-1%C3%AE-mobilization-of-p-tefb-and-basal-cdk1-drives-androgen-receptor-in-prostate-cancer
#16
Xiaming Liu, Yanfei Gao, HuiHui Ye, Sean Gerrin, Fen Ma, Yiming Wu, Tengfei Zhang, Joshua Russo, Changmeng Cai, Xin Yuan, Jihong Liu, Shaoyong Chen, Steven P Balk
P-TEFb (CDK9/cyclin T) plays a central role in androgen receptor (AR)-mediated transactivation by phosphorylating both RNA polymerase 2 complex proteins and AR at S81. CDK9 dephosphorylation mobilizes P-TEFb from an inhibitory 7SK ribonucleoprotein complex, but mechanisms targeting phosphatases to P-TEFb are unclear. We show that AR recruits protein phosphatase 1α (PP1α), resulting in P-TEFb mobilization and CDK9-mediated AR S81 phosphorylation. This increased pS81 enhances p300 recruitment, histone acetylation, BRD4 binding and subsequent further recruitment of P-TEFb, generating a positive feedback loop that sustains transcription...
January 6, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28061549/fast-and-simple-detection-methods-for-the-4-base-pair-deletion-of-canine-mdr1-abcb1-gene-by-pcr-and-isothermal-amplification
#17
Cathrin P Stiedl, Karin Weber
Dogs with a 4-bp deletion in the MDR1 (or ABCB1) gene show intolerance to certain drugs routinely used in veterinary medicine, such as ivermectin, vincristine, and doxorubicin. The mutation leads to a dysfunctional P-glycoprotein drug transporter, which results in drug accumulation in the brain and severe neurotoxicity. A rapid and accurate in-house test to determine the genotype of patients in cases of acute neurotoxic signs or in tumor patients is desirable. We describe a cost-effective detection method with simple technical equipment for veterinary practice...
January 1, 2017: Journal of Veterinary Diagnostic Investigation
https://www.readbyqxmd.com/read/28059137/establishment-of-an-accurate-and-fast-detection-method-using-molecular-beacons-in-loop-mediated-isothermal-amplification-assay
#18
Wei Liu, Simo Huang, Ningwei Liu, Derong Dong, Zhan Yang, Yue Tang, Wen Ma, Xiaoming He, Da Ao, Yaqing Xu, Dayang Zou, Liuyu Huang
This study established a constant-temperature fluorescence quantitative detection method, combining loop-mediated isothermal amplification (LAMP) with molecular beacons. The advantages of LAMP are its convenience and efficiency, as it does not require a thermocycler and results are easily visualized by the naked eye. However, a major disadvantage of current LAMP techniques is the use of indirect evaluation methods (e.g., electrophoresis, SYBR Green I dye, precipitation, hydroxynaphthol blue dye, the turbidimetric method, calcein/Mn(2+) dye, and the composite probe method), which cannot distinguish between the desired products and products of nonspecific amplification, thereby leading to false positives...
January 6, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28059049/evaluation-of-the-3m%C3%A2-molecular-detection-assay-mda-2-listeria-monocytogenes-for-the-detection-of-listeria-monocytogenes-in-select-foods-and-environmental-surfaces-collaborative-study-first-action-2016-08
#19
Patrick Bird, Jonathan Flannery, Erin Crowley, James Agin, David Goins, Lisa Monteroso
The 3M<sup>™</sup> Molecular Detection Assay (MDA) 2 <em>- Listeria </em>uses loop-mediated isothermal amplification and bioluminescence detection to rapidly detect <em>Listeria </em>spp. in a broad range of food types and environmental surfaces. Using an unpaired study design, the MDA 2 - <em>Listeria </em>was compared to the U.S. Department of Agriculture Food Safety and Inspection Service <em>Microbiology Laboratory Guidebook </em>Chapter 8.09 "<em>Isolation and Identification of Listeria monocytogenes from Red Meat, Poultry, and Egg Products, and Environmental Samples</em>" reference method for the detection of <em>Listeria </em>in deli turkey and raw chicken breast fillet...
November 17, 2016: Journal of AOAC International
https://www.readbyqxmd.com/read/28056092/improvement-and-evaluation-of-loop-mediated-isothermal-amplification-for-rapid-detection-of-toxoplasma-gondii-infection-in-human-blood-samples
#20
Xi-Meng Sun, Yong-Sheng Ji, Xian-Yong Liu, Mei Xiang, Guang He, Li Xie, Jing-Xia Suo, Xun Suo
Loop-mediated isothermal amplification (LAMP), an attractive DNA amplification method, was developed as a valuable tool for the rapid detection of Toxoplasma gondii. In this study, species-specific LAMP primers were designed by targeting the AF146527 sequence, which was a conserved sequence of 200- to 300-fold repetitive 529 bp fragment of T.gondii. LAMP reaction system was optimized so that it could detect the minimal DNA sample such as a single tachyzoite or 10 copies of recombinant plasmid. No cross-reactivity was found when using DNA from other parasites as templates...
2017: PloS One
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