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LAMP PCR

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https://www.readbyqxmd.com/read/28803792/rapid-and-visual-detection-of-leptospira-in-urine-by-ligb-lamp-assay-with-pre-addition-of-dye
#1
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
#2
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
#3
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
#4
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
#5
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
https://www.readbyqxmd.com/read/28720542/one-step-reverse-transcription-loop-mediated-isothermal-amplification-assay-for-detection-of-apple-chlorotic-leaf-spot-virus
#6
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/28706439/genome-wide-retinal-transcriptome-analysis-of-endotoxin-induced-uveitis-in-mice-with-next-generation-sequencing
#7
Yiguo Qiu, Peng Yu, Ru Lin, Xinyu Fu, Bingtao Hao, Bo Lei
PURPOSE: Endotoxin-induced uveitis (EIU) is a well-established mouse model for studying human acute inflammatory uveitis. The purpose of this study is to investigate the genome-wide retinal transcriptome profile of EIU. METHODS: The anterior segment of the mice was examined with a slit-lamp, and clinical scores were evaluated simultaneously. The histological changes in the posterior segment of the eyes were evaluated with hematoxylin and eosin (H&E) staining. A high throughput RNA sequencing (RNA-seq) strategy using the Illumina Hiseq 2500 platform was applied to characterize the retinal transcriptome profile from lipopolysaccharide (LPS)-treated and untreated mice...
2017: Molecular Vision
https://www.readbyqxmd.com/read/28697821/the-first-detection-of-toxoplasma-gondii-dna-in-environmental-air-samples-using-gelatine-filters-real-time-pcr-and-loop-mediated-isothermal-lamp-assays-qualitative-and-quantitative-analysis
#8
Anna Lass, Beata Szostakowska, Krzysztof Korzeniewski, Panagiotis Karanis
Toxoplasma gondii infections are acquired through the ingestion of oocysts present in the environment. However, there is no data about their occurrence in the air or about airborne transmission of these infections. In the present paper, we report on the identification of T. gondii using rapid molecular detection methods, supported by microscopic analysis, in environmental air samples. A total of 71 samples were collected, using gelatine filters, from kitchen gardens, recreational areas and sandpits located in northern and north-eastern Poland...
July 12, 2017: Parasitology
https://www.readbyqxmd.com/read/28697662/development-of-a-loop-mediated-isothermal-amplification-lamp-assay-for-the-detection-of-dickeya-spp
#9
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
#10
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/28677087/cloning-of-bbtv-banana-bunchy-top-virus-components-and-screening-of-bbtv-using-functionalized-gold-nanoparticles
#11
P Kumar, V Arun, T S Lokeswari
Banana bunchy top virus (BBTV) affects all varieties of banana plants and causes heavy economic loss in most of the banana cultivating areas. The BBTV genome comprises of six DNA components; in this study, we have cloned the six BBTV-DNA components from one of the BBTV-infected plants (Tri-8) and were submitted to GenBank. Analysis of the BBTV DNA-R component showed that it belonged to south Pacific group. Resistance against BBTV has not been observed so far in banana plants and removal and killing of the infected plants has been routinely practiced...
July 2017: 3 Biotech
https://www.readbyqxmd.com/read/28649808/development-of-a-real-time-loop-mediated-isothermal-amplification-assay-for-detection-of-burkholderia-mallei
#12
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
#13
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...
August 2017: Tropical Animal Health and Production
https://www.readbyqxmd.com/read/28628614/lamphimerus-a-novel-lamp-assay-for-detecting-amphimerus-sp-dna-in-human-stool-samples
#14
William Cevallos, Pedro Fernández-Soto, Manuel Calvopiña, Cristina Fontecha-Cuenca, Hiromu Sugiyama, Megumi Sato, Julio López Abán, Belén Vicente, Antonio Muro
BACKGROUND: Amphimeriasis is a fish-borne disease caused by the liver fluke Amphimerus spp. that has recently been reported as endemic in the tropical Pacific side of Ecuador with a high prevalence in humans and domestic animals. The diagnosis is based on the stool examination to identify parasite eggs, but it lacks sensitivity. Additionally, the morphology of the eggs may be confounded with other liver and intestinal flukes. No immunological or molecular methods have been developed to date...
June 2017: PLoS Neglected Tropical Diseases
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
#15
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
#16
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/28619012/genetic-analysis-of-varicella-zoster-virus-in-the-aqueous-humor-in-uveitis-with-severe-hyphema
#17
Mayumi Hosogai, Yoko Nakatani, Kensuke Mimura, Shoji Kishi, Hideo Akiyama
BACKGROUND: Genetic variations have been identified in the genome of varicella-zoster virus (VZV) strains using vesicle fluid, varicella scabs and throat swab samples. We report a rare case of VZV-associated uveitis with severe hyphema, which was immediately diagnosed by polymerase chain reaction (PCR) using the aqueous humor, in which we were able to analyze the VZV genotype for the first time. CASE PRESENTATION: A 16-year-old Japanese boy was referred to our hospital with a 20-day history of unilateral anterior uveitis and 11-day history of hyphema...
June 15, 2017: BMC Infectious Diseases
https://www.readbyqxmd.com/read/28591662/a-loop-mediated-isothermal-amplification-lamp-assay-for-the-rapid-detection-of-enterococcus-spp-in-water
#18
Roland Martzy, Claudia Kolm, Kurt Brunner, Robert L Mach, Rudolf Krska, Hana Šinkovec, Regina Sommer, Andreas H Farnleitner, Georg H Reischer
Faecal pollution of water and the resulting potential presence of human enteric pathogens is a predominant threat to public health. Microbiological water quality can be assessed by the detection of standard faecal indicator bacteria (SFIB) such as E. coli or certain Enterococcus species. In recent years, isothermal amplification methods have become a useful alternative to polymerase chain reaction (PCR), allowing molecular diagnostics with simple or no instrumentation. In this study, a novel screening method for the molecular detection of Enterococcus spp...
May 13, 2017: Water Research
https://www.readbyqxmd.com/read/28591309/detection-and-molecular-characterisation-of-a-diagnosis-escape-variant-associated-with-occult-hepatitis-b-virus-in-brazil
#19
Ricardo Wagner de Almeida, Francisco Campello do Amaral Mello, Isabelle Vasconcelos Menegoy, Márcia Paschoal do Espírito Santo, Cléber Ferreira Ginuíno, Paulo Sérgio Fonseca de Sousa, Livia Melo Villar, Elisabeth Lampe, Lia Laura Lewis-Ximenez
BACKGROUND: Many studies have identified mutations in the hepatitis B surface antigen (HBsAg) as important factors limiting the ability of commercial serological assays to detect this viral antigen. However, an association between mutations in the HBsAg gene and the occurrence of occult HBV infection (OBI) in patients has not been established. OBJECTIVES: To detect hepatitis B virus (HBV) DNA in patients with anti-HBc as a unique serological marker, a previously published, cost-effective TaqMan-based real-time polymerase chain reaction (PCR) test with minor groove binding probes was adapted for use in this study...
July 2017: Memórias do Instituto Oswaldo Cruz
https://www.readbyqxmd.com/read/28585379/groel-is-a-suitable-genetic-marker-for-detecting-vibrio-parahaemolyticus-by-loop-mediated-isothermal-amplification-assay
#20
M P Siddique, W J Jang, J M Lee, S H Ahn, S Suraiya, C H Kim, I S Kong
A groEL gene-based loop-mediated isothermal amplification (LAMP) assay was developed to detect Vibrio parahaemolyticus in contaminated seafood and water. The assay was optimized and conducted at 63°C for 40 min using Bacillus stearothermophilus (Bst) DNA polymerase, large fragment. Amplification was analysed via multiple detection methods, including opacity, formation of white precipitate, DNA intercalating dyes (ethidium bromide and SYBR Green I), metal ion-binding indicator dye, calcein, and 2% agarose gel electrophoresis...
June 5, 2017: Letters in Applied Microbiology
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