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LAMP PCR

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https://www.readbyqxmd.com/read/28916313/virulence-of-thermolable-haemolysi-tlh-gastroenteritis-related-pathogenicity-tdh-and-trh-of-the-pathogens-vibrio-parahemolyticus-in-viable-but-non-culturable-vbnc-state
#1
Huamin Zhong, Yukui Zhong, Qiulian Deng, Zhenwen Zhou, Xiaoshan Guan, Muxia Yan, Tingting Hu, Mingyong Luo
In the Viable but Non-Culturable (VBNC) state, microorganisms may survive under severe external environment. In this study, the specificity and sensitivity of PMA-LAMP assay on the detection of Vibrio Parahemolyticus (V. parahemolyticus) has been developed and evaluated, with further application on a number of food-borne V. parahemolyticus strains. Six primers were designed for recognizing 8 distinct targeting on tlh, tdh and trh gene. Through specific penetration through the damaged cell membrane of dead cells and intercalating into DNA, PMA could prevent DNA amplification of dead bacteria from LAMP, which enabled the differentiation of bacteria between VBNC state and dead state...
September 12, 2017: Microbial Pathogenesis
https://www.readbyqxmd.com/read/28915693/the-application-of-is6110-baced-loop-mediated-isothermal-amplification-lamp-in-the-early-diagnosis-of-tuberculous-meningitis
#2
Wen-Wen Sun, Qin Sun, Li-Ping Yan, Qing Zhang
Here, we evaluated the potential activity of rapid Mycobacterium tuberculosis detection with loop-mediated isothermal amplification (LAMP), for the early diagnosis of tuberculous meningitis (TBM). Patients with suspected TBM from January 2014 to December 2015 were reviewed retrospectively. The cerebrospinalfluid(CSF) was collected. Acid-fast bacillus (AFB) staining, MGIT 960 culture, real-time fluorescent quantitative polymerase chain reaction (RTFQ PCR) and LAMP were performed. A total of 200 patients were included in the study...
August 22, 2017: Oncotarget
https://www.readbyqxmd.com/read/28903726/a-novel-diagnostic-method-for-malaria-using-loop-mediated-isothermal-amplification-lamp-and-minion%C3%A2-nanopore-sequencer
#3
Kazuo Imai, Norihito Tarumoto, Kazuhisa Misawa, Lucky Ronald Runtuwene, Jun Sakai, Kyoko Hayashida, Yuki Eshita, Ryuichiro Maeda, Josef Tuda, Takashi Murakami, Shigefumi Maesaki, Yutaka Suzuki, Junya Yamagishi, Takuya Maeda
BACKGROUND: A simple and accurate molecular diagnostic method for malaria is urgently needed due to the limitations of conventional microscopic examination. In this study, we demonstrate a new diagnostic procedure for human malaria using loop mediated isothermal amplification (LAMP) and the MinION™ nanopore sequencer. METHODS: We generated specific LAMP primers targeting the 18S-rRNA gene of all five human Plasmodium species including two P. ovale subspecies (P...
September 13, 2017: BMC Infectious Diseases
https://www.readbyqxmd.com/read/28889066/effect-of-lentivirus-mediated-gene-silencing-targeting-toll-like-receptor-2-on-corneal-allograft-transplantation-in-rats
#4
Lang Bai, Weiyi Liang, Minting Chen, Yacouba Cissé, Jing Liu, Yaru Su, Jian Yu, Qiong Liu
AIM: The present work aims to assess the effectiveness of lentiviral vector (LV) mediated Toll-like receptor 2 (TLR2) gene silencing in the survival of transplanted corneal allografts, against immune rejection, in rats. METHODS: LV mediated TLR2 small interference RNA (SiRNA) with enhanced green fluorescent protein (eGFP) [LV-TLR2-siRNA-eGFP] was realised and transfected to both rat corneal epithelial (EC) and stromal cells (SC). Multiplicity of infection (MOI) was optimized for transfection efficiency using flow cytometric (FCM) analysis...
September 7, 2017: Molecular Immunology
https://www.readbyqxmd.com/read/28888887/the-novel-loop-mediated-isothermal-amplification-based-confirmation-methodology-on-the-bacteria-in-viable-but-non-culturable-vbnc-state
#5
REVIEW
Yanmei Li, Ling Yang, Jie Fu, Muxia Yan, Dingqiang Chen, Li Zhang
As a self-protection mechanism, the viable but non-culturable (VBNC) state provides the ability against conventional detection methods among various foodborne pathogens. The ability of forming colonies is lost while metabolism is still maintaining in VBNC state cells. Recently, ethidium monoazide (EMA) and propidium monoazide (PMA) have been widely applied on the detection of foodborne pathogens in VBNC state. Combined with loop-mediated isothermal amplification (LAMP), the PMA/EMA-LAMP showed a significant priority in high sensitivity, specificity and rapidity over conventional PCR based assays...
September 6, 2017: Microbial Pathogenesis
https://www.readbyqxmd.com/read/28888778/mincle-inhibits-neutrophils-and-macrophages-apoptosis-in-a-fumigatus-keratitis
#6
Jing Lin, Kun He, Guiqiu Zhao, Cui Li, Liting Hu, Guoqiang Zhu, Yawen Niu, Guipei Hao
PURPOSE: To determine whether macrophage inducible C-type lectin (Mincle) regulates neutrophils and macrophages apoptosis in A. fumigatus keratitis. MATERIALS AND METHODS: A murine model (C57BL/6) of fungal keratitis (AF) was established by gently scraping corneal central epithelium, smearing A. fumigatus on the epithelium surface and covering the eye with contact lenses. AF cell model was established by extracting neutrophils (PMN) and macrophages, and then infecting cells with A...
September 7, 2017: International Immunopharmacology
https://www.readbyqxmd.com/read/28887007/yersinia-pestis-detection-by-loop-mediated-isothermal-amplification-combined-with-magnetic-bead-capture-of-dna
#7
Na Feng, Yazhou Zhou, Yanxiao Fan, Yujing Bi, Ruifu Yang, Yusen Zhou, Xiaoyi Wang
We developed a loop-mediated isothermal amplification (LAMP) assay for the detection of Y. pestis by targeting the 3a sequence on chromosome. All 11 species of the genus Yersinia were used to evaluate the specificity of LAMP and PCR, demonstrating that the primers had a high level of specificity. The sensitivity of LAMP or PCR was 2.3 or 23CFU for pure culture, whereas 2.3×10(4) or 2.3×10(6)CFU for simulated spleen and lung samples. For simulated liver samples, the sensitivity of LAMP was 2.3×10(6)CFU, but PCR was negative at the level of 2...
August 26, 2017: Brazilian Journal of Microbiology: [publication of the Brazilian Society for Microbiology]
https://www.readbyqxmd.com/read/28886202/fuchs-endothelial-corneal-dystrophy-and-rna-foci-in-patients-with-myotonic-dystrophy
#8
V Vinod Mootha, Brock Hansen, Ziye Rong, Pradeep P Mammen, Zhengyang Zhou, Chao Xing, Xin Gong
Purpose: The most common cause of Fuchs' endothelial corneal dystrophy (FECD) is an intronic CTG repeat expansion in TCF4. Expanded CUG repeat RNA colocalize with splicing factor, muscleblind-like 1 (MBNL1), in nuclear foci in endothelium as a molecular hallmark. Myotonic dystrophy type 1 (DM1) is a neuromuscular disorder caused by a CTG repeat expansion in the 3'-untranslated region (UTR) of DMPK. In this study, we examine for RNA-MBNL1 foci in endothelial cells of FECD subjects with DM1, test the hypothesis that DM1 patients are at risk for FECD, and determine prevalence of TCF4 and DMPK expansions in a FECD cohort...
September 1, 2017: Investigative Ophthalmology & Visual Science
https://www.readbyqxmd.com/read/28866364/development-of-rapid-and-highly-sensitive-detection-of-bean-common-mosaic-necrosis-virus-in-leguminous-crops-using-loop-mediated-isothermal-amplification-assay
#9
Siwon Lee, Heejung Kim, Jin-Young Lee, Jae-Young Rho
Bean common mosaic necrosis virus (BCMNV) is a plant pathogenic virus that can infect leguminous crops such as kidney beans, sunn hemp, red beans, and mung beans. BCMNV has not been reported in Korea and is classified as a quarantine plant virus. Currently, the standard diagnostic method for diagnosis of BCMNV is reverse transcription (RT)-nested PCR system. However a more rapid monitoring system is needed to enable the testing of more samples. The use of highly efficient loop-mediated isothermal amplification (LAMP) assay for its detection has not yet been reported, and development of LAMP for detecting BCMNV in this study...
September 1, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28854526/a-comparative-study-of-three-different-gene-expression-analysis-methods
#10
Jae Young Choe, Hyung Soo Han, Seon Duk Lee, Hanna Lee, Dong Eun Lee, Jae Yun Ahn, Hyun Wook Ryoo, Jong Kun Kim
BACKGROUND: TNF-α regulates immune cells and acts as an endogenous pyrogen. Reverse transcription polymerase chain reaction (RT-PCR) is one of the most commonly used methods for gene expression analysis. Among the alternatives to PCR, loop-mediated isothermal amplification (LAMP) shows good potential in terms of specificity and sensitivity. OBJECTIVE: However, few studies have compared RT-PCR and LAMP for human gene expression analysis. Therefore, in the present study, we compared one-step RT-PCR, two-step RT-LAMP and one-step RT-LAMP for human gene expression analysis...
August 18, 2017: Technology and Health Care: Official Journal of the European Society for Engineering and Medicine
https://www.readbyqxmd.com/read/28851372/diagnostic-testing-for-legionnaires-disease
#11
REVIEW
David M Pierre, Julianne Baron, Victor L Yu, Janet E Stout
Legionnaires' disease is commonly diagnosed clinically using a urinary antigen test. The urinary antigen test is highly accurate for L. pneumophila serogroup 1, however other diagnostic tests should also be utilized in conjunction with the urinary antigen as many other Legionella species and serogroups are pathogenic. Culturing of patient specimens remains the gold standard for diagnosis of Legionnaires' disease. Selective media, BYCE with the addition of antibiotics, allows for a high sensitivity and specificity...
August 29, 2017: Annals of Clinical Microbiology and Antimicrobials
https://www.readbyqxmd.com/read/28840685/-application-of-rapid-pcr-to-authenticate-ranae-oviductus
#12
Zhi-Yi He, Xian-Ming Tang, Jian-Hui Liu, Yuan Yuan, Chao Jiang, Yu-Yang Zhao, Yang Wang
Rapid allele-specific PCR primer was designed base on Cytb 155 A/T single nucleotide polymorphism, DNA was extracted by alkaline lysis and the PCR reaction systems including denatured and annealing temperature and cycle numbers were optimized. The results were performed to authenticate Ranae Oviductus and its 4 adulterants. When 100×SYBR Green I was added in the PCR product at 90 ℃ denatured 3 s, 62 ℃ annealing 20 s and 32 cycle. Ranae Oviductus visualized strong green fluorescence under 365 nm UV lamp whereas adulterants appeared negative...
July 2017: Zhongguo Zhong Yao za Zhi, Zhongguo Zhongyao Zazhi, China Journal of Chinese Materia Medica
https://www.readbyqxmd.com/read/28837842/defining-the-relative-performance-of-isothermal-assays-that-can-be-used-for-rapid-and-sensitive-detection-of-foot-and-mouth-disease-virus
#13
Emma L A Howson, Yohei Kurosaki, Jiro Yasuda, Masayoshi Takahashi, Hiroaki Goto, Ashley R Gray, Valerie Mioulet, Donald P King, Veronica L Fowler
This study describes the first multiway comparison of portable isothermal assays for the detection of foot-and-mouth disease virus (FMDV), benchmarked against real-time reverse transcription RT-PCR (rRT-PCR). The selected isothermal chemistries included reverse transcription loop-mediated isothermal amplification (RT-LAMP) and reverse transcription recombinase polymerase amplification (RT-RPA). The analytical sensitivity of RT-LAMP was comparable to rRT-PCR (10(1) RNA copies), while RT-RPA was one log10 less sensitive (10(2) RNA copies)...
August 31, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28835287/development-of-a-loop-mediated-isothermal-amplification-technique-and-comparison-with-quantitative-real-time-pcr-for-the-rapid-visual-detection-of-canine-neosporosis
#14
Aongart Mahittikorn, Nipa Thammasonthijarern, Amonrattana Roobthaisong, Ruenruetai Udonsom, Supaluk Popruk, Sukhontha Siri, Hirotake Mori, Yaowalark Sukthana
BACKGROUND: Dogs are the definitive hosts of Neospora caninum and play an important role in the transmission of the parasite. Despite the high sensitivity of existing molecular tools such as quantitative real-time PCR (qPCR), these techniques are not suitable for use in many countries because of equipment costs and difficulties in implementing them for field diagnostics. Therefore, we developed a simplified technique, loop-mediated isothermal amplification (LAMP), for the rapid visual detection of N...
August 23, 2017: Parasites & Vectors
https://www.readbyqxmd.com/read/28826997/detection-of-skeletonema-costatum-based-on-loop-mediated-isothermal-ampli%C3%AF-cation-combined-with-lateral-%C3%AF-ow-dipstick
#15
Hai-Long Huang, Peng Zhu, Cheng-Xu Zhou, Xiao-Jun Yan, Yi-Xin Zou, Pei-Wen Lv
We developed a new assay method, which combines loop-mediated isothermal amplification (LAMP) with a chromatographic lateral flow dipstick (LFD) for the rapid and special detection of the diatom Skeletonema costatum. Four groups of LAMP primers were derived from a conserved DNA sequence unique to S. costatum. The amplifications were carried out at 61, 63, and 65 °C for 60 min in various combinations by the quantitative PCR thermal cycler to confirm optimal primers and reaction temperature. The LAMP-LFD detection limit was 0...
August 18, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28803792/rapid-and-visual-detection-of-leptospira-in-urine-by-ligb-lamp-assay-with-pre-addition-of-dye
#16
Syed Atif Ali, Gurpreet Kaur, Nongthombam Boby, T Sabarinath, Khushal Solanki, Dheeraj Pal, Pallab Chaudhuri
Leptospirosis is considered to be the most widespread zoonotic disease caused by pathogenic species of Leptospira. The present study reports a novel set of primers targeting LigB gene for visual detection of pathogenic Leptospira in urine samples through Loop-mediated isothermal amplification (LAMP). The results were recorded by using Hydroxyl napthol blue (HNB), SYBR GREEN I and calcein. Analytical sensitivity of LAMP was as few as 10 leptospiral organisms in spiked urine samples from cattle and dog. LigB gene based LAMP, termed as LigB-LAMP, was found 10 times more sensitive than conventional PCR...
August 10, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28796820/identification-of-plasmodium-spp-in-neotropical-primates-of-maranhense-amazon-in-northeast-brazil
#17
Mayra Araguaia Pereira Figueiredo, Silvia Maria Di Santi, Wilson Gómez Manrique, Marcos Rogério André, Rosangela Zacarias Machado
In the Brazilian Amazon region, malaria caused by Plasmodium malariae is considered to be a zoonosis because of cross-transfer of the parasite between humans and Neotropical primates. To contribute information on this issue, we investigated occurrences of natural infection with Plasmodium sp. among Neotropical primates in the Maranhense Amazon (Amazon region of the state of Maranhão), in the northeastern region of Brazil. Blood samples were collected from 161 Neotropical primates of six species that were caught in an environmental reserve (Sítio Aguahy) and from captive primates (CETAS-Wildlife Screening Center, municipality of São Luís), in Maranhão...
2017: PloS One
https://www.readbyqxmd.com/read/28780114/development-of-a-novel-loop-mediated-isothermal-amplification-assay-for-the-detection-of-lipolytic-pseudomonas-fluorescens-in-raw-cow-milk-from-north-china
#18
Liang Xin, Lanwei Zhang, Zhaoxu Meng, Kai Lin, Shuang Zhang, Xue Han, HuaXi Yi, Yanhua Cui
Lipases secreted by psychrotrophic bacteria are known to be heat resistant and can remain active even after the thermal processing of milk products. Such enzymes are able to destabilize the quality of milk products by causing a rancid flavor. Rapid detection of a small amount of heat-resistant lipase-producing psychrotrophic bacteria is crucial for reducing their adverse effects on milk quality. In this study, we established and optimized a novel loop-mediated isothermal amplification (LAMP) assay for the detection of Pseudomonas fluorescens in raw cow milk, as the most frequently reported heat-resistant lipase-producing bacterial species...
August 2, 2017: Journal of Dairy Science
https://www.readbyqxmd.com/read/28768730/detection-of-plasmodium-spp-infection-by-the-illumigene%C3%A2-malaria-assay-compared-to-reference-microscopy-and-real-time-pcr
#19
Candace Rypien, Barbara Chow, Wilson Chan, Deirdre Church, Dylan R Pillai
Malaria is one of the leading causes of infectious disease in returning travelers from the tropics. Diagnosis of malaria is typically performed by examination of Giemsa-stained thick and thin peripheral blood smears, which is time-consuming, labor-intensive and requires high levels of proficiency. Alternatively, loop-mediated isothermal amplification (LAMP) is a new molecular method, which is rapid, sensitive, and requires less capital equipment and technological training. We conducted a retrospective study comparing two formats of a commercial LAMP assay (Meridian illumigene® Malaria [M] and Malaria Plus [MP]) versus reference microscopy on archived blood specimens (n=140) obtained from unique returning travelers suspected of having malaria...
August 2, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28756184/development-and-validation-of-a-new-loop-mediated-isothermal-amplification-for-detection-of-pathogenic-leptospira-species-in-clinical-materials
#20
Yun-Hsiu Hsu, Shih-Jen Chou, Chih-Cheng Chang, Ming-Jeng Pan, Wei-Cheng Yang, Chuen-Fu Lin, Kun-Wei Chan
Leptospirosis, a global zoonotic disease, is often neglected but has a significant impact on human health. Here we reported a new loop mediated isothermal amplification (LAMP) assay targeting lipL32 gene of pathogenic Leptospira spp. Polymerase chain reaction (PCR), nested-PCR and real-time PCR assays were included in this study for the comparison of analytic and diagnostic sensitivity and specificity. LipL32 LAMP we designed enables detection of 10 copies of L. interrogans and has a higher diagnostic sensitivity (91...
July 26, 2017: Journal of Microbiological Methods
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