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https://www.readbyqxmd.com/read/28649808/development-of-a-real-time-loop-mediated-isothermal-amplification-assay-for-detection-of-burkholderia-mallei
#1
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
#2
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/28628614/lamphimerus-a-novel-lamp-assay-for-detecting-amphimerus-sp-dna-in-human-stool-samples
#3
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 19, 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
#4
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
#5
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
#6
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
#7
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
#8
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-lamp-assay
#9
Mahbubul Pratik Siddique, Won Je Jang, Jong Min Lee, Sun-Hee Ahn, Sharmin Suraiya, Chang-Hoon Kim, In-Soo 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 analyzed 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
https://www.readbyqxmd.com/read/28585170/rapid-and-simple-identification-of-carbapenemase-genes-bla-ndm-bla-oxa-48-bla-vim-bla-imp-14-and-bla-kpc-groups-in-gram-negative-bacilli-by-in-house-loop-mediated-isothermal-amplification-with-hydroxynaphthol-blue-dye
#10
Arpasiri Srisrattakarn, Aroonlug Lulitanond, Chotechana Wilailuckana, Nicha Charoensri, Lumyai Wonglakorn, Pimjai Saenjamla, Prajuab Chaimanee, Jureerut Daduang, Aroonwadee Chanawong
Carbapenem-resistant Enterobacteriaceae isolates by carbapenemase production are being reported globally with increasing frequency, leading to limited therapeutic options. We therefore developed a loop-mediated isothermal amplification method with hydroxynaphthol blue dye (LAMP-HNB) for rapid confirmation of bla NDM, bla OXA-48, bla VIM, bla IMP-14 and bla KPC groups. Sixty-two Enterobacteriaceae and Pseudomonas spp. isolates carrying various carbapenemase genes (28 bla NDM-1, 9 bla IMP-14a, 2 bla IMP-48, 1 bla IMP-1, 1 bla IMP-4, 1 bla IMP-9, 1 bla IMP-15, 4 bla VIM-2, 1 bla VIM-1, 1 bla IMP-14a & bla VIM-2, 7 bla KPC-2, 3 bla OXA-48 and 3 bla OXA-181) and 37 non-carbapenemase-producing Enterobacteriaceae isolates as confirmed by the PCR methods were included...
July 2017: World Journal of Microbiology & Biotechnology
https://www.readbyqxmd.com/read/28577493/inhibition-of-pi3k-akt-mtor-signaling-pathway-protects-against-d-galactosamine-lipopolysaccharide-induced-acute-liver-failure-by-chaperone-mediated-autophagy-in-rats
#11
Yin Li, Ling Lu, Ning Luo, Yong-Qiang Wang, Hong-Mei Gao
OBJECTIVE: This study aims to investigate the effects of PI3K/AKt/mTOR signaling pathway on the proliferation and apoptosis in acute liver failure (ALF) by chaperone mediated autophagy (CMA). METHODS: The hepatocytes extracted from both normal rats and rats with ALF were assigned to control, acute injury, P13K agonist, and P13K inhibitor groups. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting were used as part of this investigation to detect the expression of PI3K/AKt/mTOR signaling pathway related-proteins (PI3K, AKt, mTOR), apoptosis related-proteins (Fas, Bax, Bcl-2), chaperone-mediated autophagy (CMA) marker proteins (LAMP-2A, HSC 70), p-PI3K, p-AKt, p-4E-BPI, and p-S6K...
August 2017: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
https://www.readbyqxmd.com/read/28573341/identification-of-methicillin-resistant-staphylococcus-aureus-mrsa-using-simultaneous-detection-of-meca-nuc-and-femb-by-loop-mediated-isothermal-amplification-lamp
#12
Changguo Chen, Qiangyuan Zhao, Jianwei Guo, Yanjun Li, Qiuyuan Chen
The aim of this study was to develop a rapid detection assay to identify methicillin-resistant Staphylococcus aureus by simultaneous testing for the mecA, nuc, and femB genes using the loop-mediated isothermal amplification (LAMP) method. LAMP primers were designed using online bio-software ( http://primerexplorer.jp/e/ ), and amplification reactions were performed in an isothermal temperature bath. The products were then examined using 2% agarose gel electrophoresis. MecA, nuc, and femB were confirmed by triplex TaqMan real-time PCR...
June 2, 2017: Current Microbiology
https://www.readbyqxmd.com/read/28551733/rt-lamp-assay-an-alternative-approach-for-profiling-of-bovine-heat-shock-protein-70-gene-in-pbmc-cultured-model
#13
Gyanendra Singh Sengar, Rajib Deb, T V Raja, Umesh Singh, Rajiv Kant, V Bhanuprakash, R R Alyethodi, Sushil Kumar, Preetam Verma, Soumendu Chakraborty, Rani Alex, Rani Singh
The purpose of this study is to develop a novel Reverse Transcriptase Loop-mediated isothermal amplification (RT-LAMP) based assay for in vitro profiling of heat shock protein 70 (Hsp70) in bovine peripheral blood mononuclear cell (PBMC) culture model utilizing the absorbance level of magnesium pyrophosphate-a by-product of LAMP reaction. A set of bovine Hsp70 specific RT-LAMP primers were designed to detect the differential absorbance level of magnesium pyrophosphate by-product which signifies the degree of Hsp70 amplification from cDNA of thermally induced cultured cells at different recovery periods...
May 27, 2017: Molecular Biology Reports
https://www.readbyqxmd.com/read/28544047/comparison-of-oral-fluid-collection-methods-for-the-molecular-detection-of-hepatitis-b-virus
#14
Moyra Machado Portilho, Ana Carolina da Fonseca Mendonça, Vanessa Alves Marques, Leticia Cancella Nabuco, Cristiane Alves Villela-Nogueira, Claudia Ivantes, Lia Laura Lewis-Ximenez, Elisabeth Lampe, Livia Melo VIllar
BACKGROUND: This study aims to compare the efficiency of four oral fluid collection methods (Salivette, FTA Card, spitting and DNA-Sal) to detect HBV DNA by qualitative PCR. MATERIALS AND METHODS: Seventy-four individuals (32 HBV reactive and 42 with no HBV markers) donated serum and oral fluid. In-house qualitative PCR to detect HBV was used for both samples and commercial quantitative PCR for serum. RESULTS: HBV DNA was detected in all serum samples from HBV-infected individuals, and it was not detected in control group...
May 19, 2017: Oral Diseases
https://www.readbyqxmd.com/read/28542573/development-of-real-time-pcr-and-loop-mediated-isothermal-amplification-lamp-assays-for-the-differential-detection-of-digital-dermatitis-associated-treponemes
#15
Kelly Anklam, Megan Kulow, Wataru Yamazaki, Dörte Döpfer
Bovine digital dermatitis (DD) is a severe infectious cause of lameness in cattle worldwide, with important economic and welfare consequences. There are three treponeme phylogroups (T. pedis, T. phagedenis, and T. medium) that are implicated in playing an important causative role in DD. This study was conducted to develop real-time PCR and loop-mediated isothermal amplification (LAMP) assays for the detection and differentiation of the three treponeme phylogroups associated with DD. The real-time PCR treponeme phylogroup assays targeted the 16S-23S rDNA intergenic space (ITS) for T...
2017: PloS One
https://www.readbyqxmd.com/read/28540698/introduction-on-using-the-fastpcr-software-and-the-related-java-web-tools-for-pcr-and-oligonucleotide-assembly-and-analysis
#16
Ruslan Kalendar, Timofey V Tselykh, Bekbolat Khassenov, Erlan M Ramanculov
This chapter introduces the FastPCR software as an integrated tool environment for PCR primer and probe design, which predicts properties of oligonucleotides based on experimental studies of the PCR efficiency. The software provides comprehensive facilities for designing primers for most PCR applications and their combinations. These include the standard PCR as well as the multiplex, long-distance, inverse, real-time, group-specific, unique, overlap extension PCR for multi-fragments assembling cloning and loop-mediated isothermal amplification (LAMP)...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28534230/trueperella-pyogenes-isolated-from-a-brain-abscess-of-an-adult-roebuck-capreolus-capreolus
#17
Jörn-Peter Wickhorst, Abdulwahed Ahmed Hassan, Omar Hashim Sheet, Tobias Eisenberg, Osama Sammra, Mazen Alssahen, Christoph Lämmler, Ellen Prenger-Berninghoff, Michael Zschöck, Markus Timke, Amir Abdulmawjood
The present study was designed to characterize phenotypically and genotypically a Trueperella pyogenes strain isolated from a brain abscess of an adult roebuck (Capreolus capreolus). The species identity could be confirmed by phenotypical investigations, by MALDI-TOF MS analysis, and by sequencing the 16S ribosomal RNA (rRNA) gene, the 16S-23S rRNA intergenic spacer region (ISR); by sequencing the target genes rpoB, gap, and tuf; and by detection of T. pyogenes chaperonin-encoding gene cpn60 with a previously developed loop-mediated isothermal amplification (LAMP) assay...
May 23, 2017: Folia Microbiologica
https://www.readbyqxmd.com/read/28493440/tear-production-intraocular-pressure-and-conjunctival-bacterial-flora-in-selected-captive-wild-ruminants
#18
Pavel Kvapil, Tina Pirš, Brigita Slavec, Roman Luštrik, Tadej Zemljič, Eva Bártová, Bojana Stranjac, Marjan Kastelic
OBJECTIVE: Evaluation of tear production (Schirmer's tear test, STT) and measurement of intraocular pressure (IOP) were performed in a population of captive wild ungulates in a Slovenian ZOO during routine annual health check. ANIMALS STUDIED: In total, 10 fallow deer (Dama dama), 25 mouflons (Ovis aries musimon), 20 alpine ibexes (Capra ibex), and three alpine chamois (Rupicapra rupicapra) were included in the study. METHODS: Tear production was performed by Schirmer's tear test, IOP was measured with an applanation tonometer, and ophthalmological examination was conducted with slit-lamp biomicroscopy and indirect ophthalmoscopy...
May 11, 2017: Veterinary Ophthalmology
https://www.readbyqxmd.com/read/28493052/development-of-a-colorimetric-reverse-transcription-loop-mediated-isothermal-amplification-assay-for-the-detection-of-mirafiori-lettuce-big-vein-virus
#19
Mohammad Amin Almasi
An RT-LAMP assay was developed to detect Mirafiori lettuce big vein virus (MiLBVV) and was compared with DAS-ELISA and RT-PCR. All primers were designed on the basis of the coat protein gene of the virus. In addition, a novel immunocapture (IC) RT-LAMP assay for rapid and easy detection of MiLBVV was developed, and factors such as safety, simplicity, cost, user-friendliness and safety were compared with those of DAS-ELISA, RT-PCR and RT-LAMP assays. Compared with DAS-ELISA and RT-PCR, RT-LAMP and IC-RT-LAMP had higher sensitivity (100-fold) but similar specificity, with the advantage of a shorter assay time and no need for RNA extraction (in IC-RT-LAMP)...
May 10, 2017: Archives of Virology
https://www.readbyqxmd.com/read/28475701/crispr-cas9-mediated-mutation-of-%C3%AE-a-crystallin-gene-induces-congenital-cataracts-in-rabbits
#20
Lin Yuan, Haobin Yao, Yuxin Xu, Mao Chen, Jichao Deng, Yuning Song, Tingting Sui, Yong Wang, Yongye Huang, Zhanjun Li, Liangxue Lai
Purpose: The present study aimed to investigate the role of the αA-crystallin gene in inducing congenital cataracts in rabbits and to construct a novel animal model for characterization and pathologic analysis of congenital cataracts for future research. Methods: We generated αA-crystallin gene knockout rabbits with congenital cataracts by coinjection of Cas9 mRNA and sgRNA into zygotes. Cataract phenotypes were investigated in a repeated study of 19 F0-generation and 11 F1-generation rabbits with αA-crystallin gene mutations...
May 1, 2017: Investigative Ophthalmology & Visual Science
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