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loop-mediated isothermal amplification

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https://www.readbyqxmd.com/read/28727723/analytical-sensitivity-and-specificity-of-a-loop-mediated-isothermal-amplification-lamp-kit-prototype-for-detection-of-trypanosoma-cruzi-dna-in-human-blood-samples
#1
Susana A Besuschio, Mónica Llano Murcia, Alejandro F Benatar, Severine Monnerat, Israel Cruz Mata, Albert Picado de Puig, María de Los Ángeles Curto, Yutaka Kubota, Diana P Wehrendt, Paula Pavia, Yasuyoshi Mori, Concepción Puerta, Joseph M Ndung'u, Alejandro G Schijman
This study aimed to assess analytical parameters of a prototype LAMP kit that was designed for detection of Trypanosoma cruzi DNA in human blood. The prototype is based on the amplification of the highly repetitive satellite sequence of T.cruzi in microtubes containing dried reagents on the inside of the caps. The reaction is carried out at 65°C during 40 minutes. Calcein allows direct detection of amplified products with the naked eye. Inclusivity and selectivity were tested in purified DNA from Trypanosoma cruzi stocks belonging to the six discrete typing units (DTUs), in DNA from other protozoan parasites and in human DNA...
July 20, 2017: PLoS Neglected Tropical Diseases
https://www.readbyqxmd.com/read/28720542/one-step-reverse-transcription-loop-mediated-isothermal-amplification-assay-for-detection-of-apple-chlorotic-leaf-spot-virus
#2
Dandan Peng, Jipeng Xie, Wei Qiang, Kai-Shu Ling, Liyun Guo, Zaifeng Fan, Tao Zhou
A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the detection of Apple chlorotic leaf spot virus (ACLSV). In this method, a set of four primers was designed based on the conserved regions in the coat protein gene of ACLSV, and the primers were synthesized for the RT-LAMP assay using total RNA extracted from ACLSV-infected leaf tissues. The optimal reaction temperature and assay time were determined to be 64°C and 75min, respectively. The sensitivity of RT-LAMP reactions was reliable up to a maximum dilution of 1:3125, which was more sensitive than the RT-PCR assay...
July 15, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28716373/loop-mediated-isothermal-amplification-in-disposable-polyester-toner-microdevices
#3
Kezia Gomes de Oliveira, Juliane Cristina Borba, Alexandre Melo Bailão, Célia Maria de Almeida Soares, Emanuel Carrilho, Gabriela Rodrigues Mendes Duarte
In recent years, isothermal DNA amplification methods have emerged as an alternative in molecular diagnostics due to its ease of operation. The purpose of using isothermal amplification is to simplify the diagnostics work by i) eliminating heating cycles, ii) reducing equipment costs, and iii) providing rapid and accurate results in laboratories with limited resources. Here we show a simple and fast method for E. coli detection in disposable polyester-toner (PeT) microdevice. The amplification by LAMP of the malB gene from E...
July 14, 2017: Analytical Biochemistry
https://www.readbyqxmd.com/read/28712517/comparison-of-oropharyngeal-and-nasopharyngeal-swab-specimens-for-the-detection-of-mycoplasma-pneumoniae-in-children-with-lower-respiratory-tract-infection
#4
Fujio Kakuya, Takahiro Kinebuchi, Hitoshi Okubo, Kumihiro Matsuo
The oropharyngeal swab specimen was superior to the nasopharyngeal swab specimen for the detection of Mycoplasma pneumoniae in children with lower respiratory tract infection. The oropharyngeal loop-mediated isothermal amplification had 100% sensitivity and specificity compared with polymerase chain reaction testing, whereas the oropharyngeal rapid antigen detection test using immunochromatographic assay had relatively low sensitivity (66%) and reasonable specificity (90.7%).
July 13, 2017: Journal of Pediatrics
https://www.readbyqxmd.com/read/28702363/a-case-of-bacillus-calmette-gu%C3%A3-rin-cystitis-diagnosed-with-a-novel-loop-mediated-isothermal-amplification-method
#5
Kazuyoshi Tachi, Akinori Sato, Yuji Kouzaki, Takuya Maeda, Akihiko Kawana, Tomohiko Asano
Intravesical bacillus Calmette-Guérin (BCG) instillation is broadly used to prevent bladder cancer recurrence or to treat carcinoma in situ. BCG infection is rare but can cause serious problems because this strain has intrinsic resistance to pyrazinamide, a first-line anti-tuberculosis drug. Furthermore, there had been no specific and easy procedure accurately diagnosing BCG infection. In this case report we present the first case of BCG cystitis diagnosed with a newly developed easy-to-use diagnostic procedure using the loop-mediated isothermal amplification method...
September 2017: Urology Case Reports
https://www.readbyqxmd.com/read/28697662/development-of-a-loop-mediated-isothermal-amplification-lamp-assay-for-the-detection-of-dickeya-spp
#6
Jarred Hideki Yasuhara-Bell, Glorimar Marrero, Mohammad Arif, Asoka de Silva, Anne Alvarez
Dickeya and Pectobacterium spp. are responsible for soft rotting diseases of several plant species, some with overlapping host range. On potato, symptoms caused by these pathogens cannot be clearly differentiated. Disease results in the down-grading and rejection of potato seed, thus requiring additional phytosanitary restrictions across Northern Europe and other parts of the world. In an effort to provide a more timely and accurate diagnostic to distinguish these two groups of pathogens, a method for detecting Dickeya spp...
July 11, 2017: Phytopathology
https://www.readbyqxmd.com/read/28689247/loop-mediated-isothermal-amplification-as-a-reliable-assay-for-toxocara-canis-infection-in-pet-dogs
#7
Paria Khoshakhlagh, Adel Spotin, Mahmoud Mahami-Oskouei, Abbas Shahbazi, Maryam Ozlati
Keeping of infected dogs as pet results in the potential transmission risk factors for shedding helminthic infections such as toxocariasis. Lack of accurate identification of Toxocara canis eggs in non-dewormed infected pet dogs remains a diagnostic concern among researchers. In this study, dog owners were asked to fill up a questionnaire regarding their pets and their attitude towards the deworming regimen. One hundred faecal samples were collected from pet dogs (Northwest Iran) and were subsequently identified by the ZnSo4 flotation technique, PCR and loop-mediated isothermal amplification (LAMP) assays...
July 9, 2017: Parasitology Research
https://www.readbyqxmd.com/read/28684052/rapid-detection-of-trypanosoma-cruzi-by-colorimetric-loop-mediated-isothermal-amplification-lamp-a-potential-novel-tool-for-the-detection-of-congenital-chagas-infection
#8
Rocío Rivero, Margarita Bisio, Elsa Beatriz Velázquez, Mónica Inés Esteva, Karenina Scollo, Nicolás Leonel González, Jaime Altcheh, Andrés Mariano Ruiz
Early diagnosis of congenital Trypanosomacruzi transmission in newborns is essential because babies show high indices of cure. Conventional diagnosis is based on microscopic examination and serology. Molecular diagnosis is a promising alternative to replace conventional diagnosis, although it is not well suited for adoption in laboratories with limited resources. Isothermal DNA amplification methods have the advantage of not requiring expensive equipment. The aim of this work was to apply loop-mediated isothermal amplification (LAMP) to detect congenital infection in babies colorimetrically...
June 19, 2017: Diagnostic Microbiology and Infectious Disease
https://www.readbyqxmd.com/read/28683669/development-of-loop-mediated-isothermal-amplification-with-plasmodium-falciparum-unique-genes-for-molecular-diagnosis-of-human-malaria
#9
Yijing Zhang, Yi Yao, Weixing Du, Kai Wu, Wenyue Xu, Min Lin, Huabing Tan, Jian Li
In order to achieve better outcomes for treatment and in the prophylaxis of malaria, it is imperative to develop a sensitive, specific, and accurate assay for early diagnosis of Plasmodium falciparum infection, which is the major cause of malaria. In this study, we aimed to develop a loop-mediated isothermal amplification (LAMP) assay with P. falciparum unique genes for sensitive, specific, and accurate detection of P. falciparum infection. The unique genes of P. falciparum were randomly selected from PlasmoDB...
July 6, 2017: Pathogens and Global Health
https://www.readbyqxmd.com/read/28681828/development-and-evaluation-of-a-loop-mediated-isothermal-amplification-combined-with-au-nanoprobe-assay-for-rapid-detection-of-mycobacterium-tuberculosis
#10
Thongchai Kaewphinit, Jutturong Ckumdee, Kosum Chansiri, Somchai Santiwatanakul
Loop-mediated isothermal amplification (LAMP) has been proposed as an inexpensive and easy to perform assay for molecular diagnostics. We present a novel strategy for the detection of LAMP amplicons derived from Mycobacterium tuberculosis by the use of Au-nanoprobes. When applied to a total of 93 clinical specimens, the LAMP assay demonstrated sensitivity and specificity higher than that of polymerase chain reaction and culture. The Au-nanoprobe augmented LAMP test platform with its advantages of robust reagents and a simple colorimetric detection method can be adapted easily for the rapid detection of other infectious disease agents at a low cost...
April 2017: Indian Journal of Medical Microbiology
https://www.readbyqxmd.com/read/28670469/an-integrated-versatile-lab-on-a-chip-platform-for-the-isolation-and-nucleic-acid-based-detection-of-pathogens
#11
Natalia Sandetskaya, Doreen Moos, Harald Pötter, Stefan Seifert, Marcin Jenerowicz, Holger Becker, Christian Zilch, Dirk Kuhlmeier
AIM: Processing of the samples in molecular diagnostics is complex and labor-intensive. An integrated and automated platform for sample preparation and nucleic acid-based detection can significantly relieve this burden for the users. RESULTS: We present a prototype of a versatile and integrated platform for the detection of pathogens in various liquid media. We describe a proof-of-concept for the integrated isolation of bacteria, cell lysis with optional DNA extraction, DNA amplification and detection in two different reactions, loop-mediated isothermal amplification and PCR, on a single microfluidic platform...
June 2017: Future Science OA
https://www.readbyqxmd.com/read/28659561/detection-of-fungi-from-an-indoor-environment-using-loop-mediated-isothermal-amplification-lamp-method
#12
Takako Nakayama, Takashi Yamazaki, Ayaka Yo, Kazuya Tone, Mohamed Mahdi Alshahni, Ryuichi Fujisaki, Koichi Makimura
 Loop-mediated isothermal amplification (LAMP) is a useful DNA detection method with high specificity and sensitivity. The LAMP reaction is carried out within a short time at a constant temperature without the need for thermal cycling. We developed a LAMP primer set for detecting a wide range of fungi by aligning the sequences of the large subunit ribosomal RNA gene of Candida albicans (Ascomycota), Cryptococcus neoformans (Basidiomycota), and Mucor racemosus (Mucorales). The threshold of C. albicans rDNA as template with our LAMP primer set was in the range of 10-100 copies per a reaction...
2017: Biocontrol Science
https://www.readbyqxmd.com/read/28652587/novel-loop-mediated-isothermal-amplification-lamp-assay-with-a-universal-qprobe-can-detect-snps-determining-races-in-plant-pathogenic-fungi
#13
Yu Ayukawa, Saeri Hanyuda, Naoko Fujita, Ken Komatsu, Tsutomu Arie
Tomato wilt pathogen Fusarium oxysporum f. sp. lycopersici (Fol) is grouped into three races based on their pathogenicity to different host cultivars. Rapid detection and discrimination of Fol races in field soils is important to prevent tomato wilt disease. Although five types of point mutations in secreted in xylem 3 (SIX3) gene, which are characteristic of race 3, have been reported as a molecular marker for the race, detection of these point mutations is laborious. The aim of this study is to develop a rapid and accurate method for the detection of point mutations in SIX3 of Fol...
June 26, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28649808/development-of-a-real-time-loop-mediated-isothermal-amplification-assay-for-detection-of-burkholderia-mallei
#14
V Pal, A Saxena, S Singh, A K Goel, J S Kumar, M M Parida, G P Rai
Burkholderia mallei is the aetiological agent of glanders, a highly contagious and re-emerging zoonotic disease. Early diagnosis of glanders is critically important to ensure timely treatment with appropriate antibiotics in humans, and to prevent spread of infection in animals. Molecular detection of B. mallei has always been troublesome because of its genetic similarity with Burkholderia pseudomallei, the causative agent of melioidosis. In present investigation, a set of six B. mallei-specific primers were designed and a simple, rapid, specific and sensitive real-time loop-mediated isothermal amplification (LAMP) assay was developed for detection of B...
June 25, 2017: Transboundary and Emerging Diseases
https://www.readbyqxmd.com/read/28638960/comparative-diagnostic-evaluation-of-omp31-gene-based-taqman%C3%A2-real-time-pcr-assay-with-visual-lamp-assay-and-indirect-elisa-for-caprine-brucellosis
#15
Suman Saini, V K Gupta, K Gururaj, D D Singh, R V S Pawaiya, N K Gangwar, A K Mishra, Deepak Dwivedi, Dimple Andani, Ashok Kumar, T K Goswami
Brucellosis is one of the leading causes of abortion in domestic animals that imposes costs on both economy and society. The disease is highly zoonotic and poses risk to animal handlers due to its zoonotic nature. It causes stillbirth, loss of kids and abortion in last term of pregnancy. Reproductive damage includes infertility in does and orchitis and epididymitis in breeding bucks, which result in high financial losses to farmers and the agriculture industry as a whole. It requires highly sensitive and specific assays to diagnose the disease at field level...
June 21, 2017: Tropical Animal Health and Production
https://www.readbyqxmd.com/read/28628741/amplicon-competition-enables-end-point-quantitation-of-nucleic-acids-following-isothermal-amplification
#16
Yu Sherry Jiang, Apollo Stacy, Marvin Whiteley, Andrew D Ellington, Sanchita Bhadra
It is inherently difficult to quantitate nucleic acid analytes with most isothermal amplification assays. We have now developed loop-mediated isothermal amplification (LAMP) reactions in which competition between defined numbers of 'false' and 'true' amplicons leads to order of magnitude quantitation via a single end-point determination. These thresholded LAMP reactions were successfully used to directly and quantitatively estimate the numbers of nucleic acids in complex biospecimens, including directly from cells and in sewage, with the values obtained closely correlating with qPCR quantitations...
June 19, 2017: Chembiochem: a European Journal of Chemical Biology
https://www.readbyqxmd.com/read/28627822/development-of-a-one-step-reverse-transcription-loop-mediated-isothermal-amplification-system-for-a-highly-sensitive-detection-of-rabbit-hepatitis-e-virus
#17
Jing Wang, Hua Zhang, Shengyuan Yan, Yichao Wang, Xin Lu, Hongwei Fu
BACKGROUND: Hepatitis E virus (HEV) is a major cause of acute viral hepatitis in areas with poor sanitation. Rabbit is one of important animal reservoirs of the virus. Reverse transcription-loop-mediated isothermal amplification (RT-LAMP) system is a novel nucleic acid amplification assay, for which the specificity and sensitivity are much higher than that of conventional PCR. However, previously reported RT-LAMP system cannot detect rabbit HEV, which was identified recently and seems to be another potential source of human HEV infection...
May 1, 2017: Clinical Laboratory
https://www.readbyqxmd.com/read/28619672/point-of-care-diagnosis-of-multiple-schistosome-parasites-species-specific-dna-detection-in-urine-by-loop-mediated-isothermal-amplification-lamp
#18
Nilanjan Lodh, Kei Mikita, Kwabena M Bosompem, William K Anyan, Joseph K Quartey, Joseph Otchere, Clive J Shiff
Schistosomes are easily transmitted and high chance of repeat infection, so if control strategies based on targeted mass drug administration (MDA) are to succeed it is essential to have a test that is sensitive, accurate and simple to use. It is known and regularly demonstrated that praziquantel does not always eliminate an infection so in spite of the successes of control programs a residual of the reservoir survives to re-infect snails. The issue of diagnostic sensitivity becomes more critical in the assessment of program effectiveness...
June 12, 2017: Acta Tropica
https://www.readbyqxmd.com/read/28600772/a-closed-tube-loop-mediated-isothermal-amplification-assay-for-the-visual-endpoint-detection-of-brucella-spp-and-mycobacterium-avium-subsp-paratuberculosis
#19
Marcos D Trangoni, Andrea K Gioffré, Silvio L Cravero
LAMP (loop-mediated isothermal amplification) is an isothermal nucleic acid amplification technique that is characterized by its efficiency, rapidity, high yield of final product, robustness, sensitivity, and specificity, with the blueprint that it can be implemented in laboratories of low technological complexity. Despite the conceptual complexity underlying the mechanistic basis for the nucleic acid amplification, the technique is simple to use and the amplification and detection can be carried out in just one step...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28592104/-loop-mediated-isothermal-amplification-technique-in-the-diagnosis-of-hepatitis-b-virus-infection-a-meta-analysis
#20
H Y Zhou, C Chen, X N Li, X J Ma
Objective: To evaluate the diagnostic performance of Loop-mediated isothermal amplification (LAMP) in the diagnosis of Hepatitis B Virus (HBV) infection using Meta-analysis. Methods: Literatures about LAMP in the diagnosis of HBV throughPubMed database of the National Library of Medicine, the EMBASE database of the Dutch Medical Digest, the Cochrane Clinical Trials Database, China Science Periodical Database, CSPD and the China National Knowledge Infrastructure (CNKI) were searched from 2000 to 2016, and the Language limited to Chinese and English...
June 6, 2017: Zhonghua Yu Fang Yi Xue za Zhi [Chinese Journal of Preventive Medicine]
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