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Ole Lagatie, Michelle Merino, Linda Batsa Debrah, Alexander Y Debrah, Lieven J Stuyver
BACKGROUND: Diagnostic procedures for the diagnosis of infection with the nematode parasite Onchocerca volvulus are currently based on the microscopic detection of microfilariae in skin biopsies. Alternative approaches based on amplification of parasitic DNA in these skin biopsies are currently being explored. Mostly this is based on the detection of the O-150 repeat sequence using PCR based techniques. METHODS: An isothermal, loop-mediated amplification method has been designed using the mitochondrial O...
December 1, 2016: Parasites & Vectors
Ling Chen, Zhiyuan Jiao, Dongmei Liu, Xingliang Liu, Zihao Xia, Congliang Deng, Tao Zhou, Zaifeng Fan
Maize chlorotic mottle virus (MCMV) is spreading in many regions worldwide, causing maize lethal necrosis when co-infected with a potyvirid. In this study, one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed to detect MCMV in maize. A set of four specific primers was designed based on the conserved coat protein gene sequences of MCMV. The RT-LAMP could be completed within 60min under isothermal condition at 63°C. The sensitivity test showed that the RT-LAMP was about 10-fold more sensitive than RT-PCR and no cross-reactivity was detected with other viral pathogens infecting maize in China...
November 26, 2016: Journal of Virological Methods
Maria Immaculata Arifin, Johan Höglund, Adam Novobilský
The liver fluke, Fasciola hepatica, is one of the major parasite threats to livestock industries world-wide. In sheep and cattle, F. hepatica infection is commonly diagnosed using a range of methods. Aside from conventional coprological and serological diagnostic methods, there are also several molecular methods available based on the detection of liver fluke DNA in faeces. In this study, the outcomes of faecal egg count (FEC), serology and coproantigen ELISA (cELISA) were compared with the performance of polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) in diagnosis of F...
December 15, 2016: Veterinary Parasitology
Amir Abdoli, Abdolhossein Dalimi, Haleh Soltanghoraee, Fatemeh Ghaffarifar
Toxoplasma gondii is one of the most common zoonotic parasitic diseases in human and warm-blooded animals worldwide. Birds are one of important intermediate hosts of T. gondii. The aim of this study is molecular detection of T. gondii in the house sparrow by LAMP and PCR methods in Tehran, Iran. A total 200 sparrows were captured in different regions of Tehran. DNA was extracted from tissue samples of each sparrow. LAMP and conventional PCR assays were carried out with a set of primers to detect the 529 bp fragment of T...
December 2016: Journal of Parasitic Diseases: Official Organ of the Indian Society for Parasitology
Farhan Ahmad, Robert D Stedtfeld, Hassan Waseem, Maggie R Williams, Alison M Cupples, James M Tiedje, Syed A Hashsham
We are reporting a most probable number approach integrated to loop mediated isothermal technique (MPN-LAMP) focusing on Gram-negative Escherichia coli and Gram-positive Enterococcus faecalis bacterial cells without nucleic acids extraction. LAMP assays for uidA from E. coli and gelE from E. faecalis were successfully performed directly on cells up to single digit concentration using a commercial real time PCR instrument. Threshold time values of LAMP assays of bacterial cells, heat treated bacterial cells (95°C for 5min), and their purified genomic DNA templates were similar, implying that amplification could be achieved directly from bacterial cells at 63°C...
November 14, 2016: Journal of Microbiological Methods
Muxia Yan, Weidong Li, Zhenwen Zhou, Hongxia Peng, Ziyan Luo, Ling Xu
In this work, loop-mediated isothermal amplification based detection assay using bacterial culture and bacterial colony for various common pathogens direct detection had been established, evaluated and further applied. A total of five species of common pathogens and nine detection targets (tlh, tdh and trh for V. Parahaemolyticus, rfbE, stx1 and stx2 for E. coli, oprI for P. aeruginosa, invA for Salmonella and hylA for L. monocytogenes) were performed on bacterial culture and bacterial colony LAMP. To evaluate and optimize this assay, a total of 116 standard strains were included...
November 9, 2016: Microbial Pathogenesis
Geoff M Gurr, Anne C Johnson, Gavin J Ash, Bree A L Wilson, Mark M Ero, Carmel A Pilotti, Charles F Dewhurst, Minsheng S You
The recent discovery of Bogia coconut syndrome in Papua New Guinea (PNG) is the first report of a lethal yellowing disease (LYD) in Oceania. Numerous outbreaks of LYDs of coconut have been recorded in the Caribbean and Africa since the late Nineteenth century and have caused the death of millions of palms across several continents during the Twentieth century. Despite the severity of economic losses, it was only in the 1970s that the causes of LYDs were identified as phytoplasmas, a group of insect-transmitted bacteria associated with diseases in many other economically important crop species...
2016: Frontiers in Plant Science
Jin Zhao, Yubin Xing, Wei Liu, Wentao Ni, Chuanqi Wei, Rui Wang, Yunxi Liu, Youning Liu
Stenotrophomonas maltophilia is a common nosocomial pathogen that causes high morbidity and mortality. Because of its inherent extended antibiotic resistance, therapeutic options for S. maltophilia are limited, and sulfamethoxazole/trimethoprim (SXT) is the only first-line antimicrobial recommended. However, with the spread of dihydropteroate synthase (sul1 and sul2) genes, global emergence of SXT resistance has been reported. There is an urgent need to develop a rapid and sensitive but cost-efficient method to monitor the dissemination of sul genes...
2016: Frontiers in Microbiology
Naomi W Lucchi, Marie Gaye, Mammadou Alpha Diallo, Ira F Goldman, Dragan Ljolje, Awa Bineta Deme, Aida Badiane, Yaye Die Ndiaye, John W Barnwell, Venkatachalam Udhayakumar, Daouda Ndiaye
Isothermal nucleic acid amplification assays such as the loop mediated isothermal amplification (LAMP), are well suited for field use as they do not require thermal cyclers to amplify the DNA. To further facilitate the use of LAMP assays in remote settings, simpler sample preparation methods and lyophilized reagents are required. The performance of a commercial malaria LAMP assay (Illumigene Malaria LAMP) was evaluated using two sample preparation workflows (simple filtration prep (SFP)) and gravity-driven filtration prep (GFP)) and pre-dispensed lyophilized reagents...
November 9, 2016: Scientific Reports
Sylvie Kemleu, Dylan Guelig, Carole Eboumbou Moukoko, Estelle Essangui, Steven Diesburg, Abas Mouliom, Bernard Melingui, Jeanne Manga, Christiane Donkeu, Annie Epote, Gaëtan Texier, Paul LaBarre, Robert Burton, Lawrence Ayong
Highly sensitive and field deployable molecular diagnostic tools are critically needed for detecting submicroscopic, yet transmissible levels of malaria parasites prevalent in malaria endemic countries worldwide. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed and evaluated in comparison with thick blood smear microscopy, an antigen-based rapid diagnostic test (RDT), and an in-house RT-PCR targeting the same RT-LAMP transcript. The optimized assay detected Plasmodium falciparum infections in as little as 0...
2016: PloS One
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...
November 2, 2016: Parasitology International
Hadi Mirahmadi, Alireza Maleki, Raheleh Hasanzadeh, Mohammad Bagher Ahoo, Iraj Mobedi, Ali Rostami
This report describes a rare case of ophthalmic dirofilariasis in a 2-year-old boy with redness, irritation, pain and foreign body sensation in the right eye. Slit lamp examination demonstrated a thread-like whitish nematode in the anterior chamber of the right eye that twisted around itself. The nematode worm (35mm long and 150-200μm width) was removed surgically. The presence of the smooth cuticular surface without longitudinal ridges and the vulva showed that it could be a female Diroflariaimmitis. PCR amplification was done to verify the Diroflaria species...
November 1, 2016: Parasitology International
Reza Nabie, Adel Spotin, Soheila Rouhani
A rare case of human subconjunctival setariasis due to Setaria equina infection is reported herein. A 15-years old girl was referred with a 24h history of edema and redness in her left eye. On slit lamp examination, a thread-like cylindrical worm was moving in the subconjunctival area. The worm was extracted, stained and measured 110mm in length 510μm in width. The isolated worm was identified as adult female S. equina based on morphometric criteria. Identification of the species of the worm was confirmed using molecular methods...
October 26, 2016: Parasitology International
Xuan Wang, Fenggui Yin, Yuhai Bi, Gong Cheng, Jing Li, Lidan Hou, Yunlong Li, Baozhi Yang, Wenjun Liu, Limin Yang
BACKGROUND: Zika virus (ZIKV) is an arbovirus that recently emerged and has expanded worldwide, causing a global threat and raising international concerns. Current molecular diagnostics, e.g., real-time PCR and reverse transcription PCR (RT-PCR), are time consuming, expensive, and can only be deployed in a laboratory instead of for field diagnostics. OBJECTIVES: This study aimed to develop a one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) platform showing sensitivity, specificity, and more convenience than previous methods, being easily distributed and implemented...
October 25, 2016: Journal of Virological Methods
Sandamali A Ekanayaka, Sharon A McClellan, Ronald P Barrett, Shikhil Kharotia, Linda D Hazlett
Purpose: High mobility group box 1 (HMGB1) contributes to poor disease outcome in Pseudomonas aeruginosa keratitis. This study tests the prophylactic effect of treatment with HMGB1 inhibitors, glycyrrhizin (GLY) and its derivative, carbenoxolone (CBX), for Pseudomonas keratitis. Methods: We treated C57BL/6 (B6) mice subconjunctivally with GLY or CBX, infected with a noncytotoxic clinical isolate (KEI 1025) or a cytotoxic strain (ATCC 19660) of P. aeruginosa, and injected intraperitoneally with either agent...
October 1, 2016: Investigative Ophthalmology & Visual Science
L L Wei, X Y Zhang, L Zhang, R R Yang, H B Shi, T Wen, D X Chen, Z P Duan, F Ren
Objective: To investigate the expression and role of autophagy in the progression of acute liver failure (ALF) using the mouse model of ALF induced by D-galactosamine/LPS (D-GalN/LPS). Methods: The C57BL/6 mice were used, and intraperitoneal injection of D-galactosamine (D-GalN) and lipopolysaccharide (LPS) was performed to establish the mouse model of ALF. The mice were divided into control group and 2-, 4-, and 6-hour D-GalN/LPS-induced ALF model groups. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured to assess liver function, and the pathological changes in liver tissue were observed to evaluate the status of liver injury...
August 20, 2016: Zhonghua Gan Zang Bing za Zhi, Zhonghua Ganzangbing Zazhi, Chinese Journal of Hepatology
Zeynep Koloren, Emine Ayaz
This research was undertaken to study the molecular detection and characterization of Cryptosporidium spp. in environmental water sources at Samsun and Giresun Provinces of The Black Sea in Turkey. Two-hundred forty and one-hundred eighty environmental samples were collected from a total of twenty and twenty-five sampling sites of Giresun and Samsun Provinces. One hundred twenty untreated drinking water samples were also detected for Cryptosporidium spp. in both investigated areas. 101 (%42), 92 (%38.3) of 240 and 74 (41...
December 1, 2016: Acta Parasitologica
Huaqi Tang, Chunsheng Liu, Yanpeng Li, Xinyue Zhang, Guojie Xu
Fungi of the Alternaria genus are associated with allergic diseases, with Alternaria alternata being one of the most prevalent species. A. alternata has been frequently reported as the etiologic agent of hypersensitivity pneumonitis, allergic rhinosinusitis, bronchial asthma, and other diseases. In this study, we developed a loop-mediated isothermal amplification (LAMP) assay and a real-time PCR assay to detect low levels of A. alternata in herbal tea samples. The LAMP assay can detect as little as 3 pg/μL of A...
October 18, 2016: Journal of AOAC International
Yohei Kurosaki, Sayaka Okada, Sayuri Nakamae, Jiro Yasuda
Bovine papular stomatitis virus (BPSV) causes pustular cutaneous disease in cattle worldwide. This paper describes the development of a specific loop-mediated isothermal amplification (LAMP) assay to detect BPSV which did not cross-react with other parapoxviruses. To assess analytical sensitivity of this LAMP assay, DNA was extracted from serially diluted BPSV from which the infectious titer was determined by a novel assay based on calf kidney epithelial cells. The LAMP assay had equivalent analytical sensitivity to quantitative PCR, and could detect as few as 86 copies of viral DNA per reaction...
October 14, 2016: Journal of Virological Methods
Xiangfeng He, Fei Xue, Shufa Xu, Wenhe Wang
Lily symptomless virus (LSV) is one of the most prevalent viruses that infect lily plants worldwide. A rapid and sensitive reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for detection of LSV, using two primer pairs that specifically amplified the conserved sequence of LSV coat protein. The optimum reaction conditions were as follows: 4mM MgCl2 and 0.8M betaine with incubation at 64°C for 30min. The limit of detection of LSV from infected lily leaves was 10-fold higher for RT-LAMP than for conventional RT-PCR...
October 11, 2016: Journal of Virological Methods
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