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Sathish Kumar T, Navaneeth Krishnan A, Joseph Sahaya Rajan J, Makesh M, Jithendran K P, Alavandi S V, Vijayan K K
The emerging microsporidian parasite Enterocytozoon hepatopenaei (EHP), the causative agent of hepatopancreatic microsporidiosis, has been widely reported in shrimp-farming countries. EHP infection can be detected by light microscopy observation of spores (1.7 × 1 μm) in stained hepatopancreas (HP) tissue smears, HP tissue sections, and fecal samples. EHP can also be detected by polymerase chain reaction (PCR) targeting the small subunit (SSU) ribosomal RNA (rRNA) gene or the spore wall protein gene (SWP)...
March 17, 2018: Parasitology Research
E A R Engku Nur Syafirah, A B Nurul Najian, Phiaw Chong Foo, Mohammad Ridhuan Mohd Ali, Maizan Mohamed, Chan Yean Yean
Cholera, caused by Vibrio cholerae is a foodborne disease that frequently reported in food and water related outbreak. Rapid diagnosis of cholera infection is important to avoid potential spread of disease. Among available diagnostic platforms, loop-mediated isothermal amplification (LAMP) is regarded as a potential diagnostic tool due to its rapidity, high sensitivity and specificity and independent of sophisticated thermalcycler. However, the current LAMP often requires multiple pipetting steps, hence is susceptible to cross contamination...
March 12, 2018: Acta Tropica
Sonja Günther, Sandra Felten, Gerhard Wess, Katrin Hartmann, Karin Weber
Feline infectious peritonitis (FIP) is a fatal disease in cats worldwide. The aim of this study was to test two commercially available reaction mixtures in a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay to detect feline Coronavirus (FCoV) in body cavity effusions of cats with and without FIP, in order to minimize the time from sampling to obtaining results. RNA was extracted from body cavity effusion samples of 71 cats, including 34 samples from cats with a definitive diagnosis of FIP, and 37 samples of control cats with similar clinical signs but other confirmed diseases...
March 11, 2018: Journal of Virological Methods
Javier Gandasegui, Pedro Fernández-Soto, Antonio Muro, Constança Simões Barbosa, Fabio Lopes de Melo, Rodrigo Loyo, Elainne Christine de Souza Gomes
BACKGROUND: In Brazil, schistosomiasis is a parasitic disease of public health relevance, mainly in poor areas where Schistosoma mansoni is the only human species encountered and Biomphalaria straminea is one of the intermediate host snails. A nested-PCR based on a specific mitochondrial S. mansoni minisatellite DNA region has been successfully developed and applied as a reference method in Brazil for S. mansoni detection, mainly in host snails for epidemiological studies. The amplification efficiency of LAMP is known to be higher than PCR...
March 2018: PLoS Neglected Tropical Diseases
Maria Cryskely Agra Batinga, Julia Teresa Ribeiro de Lima, Fabio Gregori, Jaqueline Assumpção Diniz, Kerstin Muner, Trícia M F S Oliveira, Helena Lage Ferreira, Rodrigo Martins Soares, Lara Borges Keid
Canine brucellosis is caused by Brucella canis, a gram negative and facultative intracellular bacterium that is commonly associated with reproductive failures in dogs. The accurate diagnosis of the infection relies on the use of serological tests associated with blood culturing to guarantee sensitivity. The polymerase chain reaction (PCR) can replace the culturing procedure for the direct diagnosis of the infection because of its speed, high specificity and sensitivity values; however, it depends on some laboratory infrastructure to be conducted...
March 7, 2018: Molecular and Cellular Probes
Waqas Ali, Mudasser Habib, Rai Shafqat Ali Khan, Muhammad Ashir Zia, Muhammad Farooq, Sanaullah Sajid, Muhammad Salah Ud Din Shah
This study reports the molecular characterization of foot-and-mouth disease virus (FMDV) in the provinces of Punjab and Sindh, Pakistan during 2014-17. FMDV genome was detected in 42 and 41 out of 46 samples (epithelial tissue and saliva) by reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) and reverse transcriptase polymerase chain reaction (RT-PCR), respectively. Sequences of the complete VP1 coding region of the samples (n = 33) was achieved showing that 10, 4 and 19 samples belonged to serotype O, A and Asia1 respectively...
March 7, 2018: Archives of Virology
Moyra Machado Portilho, Ana Carolina da Fonseca Mendonça, Cristianne Bezerra da Silva, Márcia Paschoal do Espirito-Santo, Vanessa Salete de Paula, Leticia Cancella Nabuco, Cristiane Alves Villela-Nogueira, Lia Laura Lewis-Ximenez, Elisabeth Lampe, Livia Melo Villar
For quantification of hepatitis B virus DNA (HBV DNA), commercial assays are used with serum or plasma samples, but oral fluid samples could be an alternative for HBV diagnosis due to ease of collection. This study aims to develop in-house real time PCR using synthetic curve for HBV DNA quantification for serum and oral fluid samples. Samples were collected from 103 individuals (55 HBsAg reactive and HBV DNA reactive by commercial assay and 48 without HBV markers) and submitted to two in-house real time PCR assays for HBV pre-S/S region with different standard curves: qPCR plasmidial and qPCR synthetic...
March 4, 2018: Journal of Virological Methods
S Zheng, X Wu, J Shi, Z Peng, M Gao, C Xin, Y Liu, S Wang, S Xu, H Han, J Yu, W Sun, X Cong, J Li, J Wang
In this study, a rapid and specific assay for the detection of porcine circovirus type 3 (PCV3) was established using loop-mediated isothermal amplification (LAMP). Four primers were specifically designed to amplify PCV3. The LAMP assay was effectively optimized to amplify PCV3 by water bath at 60°C for 60 min. The detection limit was approximately 1 × 101 copy in this LAMP assay. Compared to porcine circovirus type 2 (PCV2), both gE and gD genes of pseudorabies virus (PRV) and porcine parvovirus (PPV), the LAMP assay showed a high specific detection of PCV3...
March 4, 2018: Transboundary and Emerging Diseases
Laura E Lamb, Sarah N Bartolone, Maya O Tree, Michael J Conway, Julien Rossignol, Christopher P Smith, Michael B Chancellor
Infection with Zika virus (ZIKV) is of growing concern since infection is associated with the development of congenital neurological disease. Quantitative reverse transcription PCR (qRT-PCR) has been the standard for ZIKV detection; however, Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) may allow for faster and cheaper testing. Studies have suggested that ZIKV detection in urine is more sensitive and has a longer window of detection compared to serum and saliva. The objective of this study was to develop a urine diagnostic test that could be completed in under 30 minutes...
February 28, 2018: Scientific Reports
Suneel Gupta, Michael K Fink, Arkasubhra Ghosh, Ratnakar Tripathi, Prashant R Sinha, Ajay Sharma, Nathan P Hesemann, Shyam S Chaurasia, Elizabeth A Giuliano, Rajiv R Mohan
Purpose: We tested the potential of bone morphogenic protein 7 (BMP7) and hepatocyte growth factor (HGF) combination gene therapy to treat preformed corneal fibrosis using established rabbit in vivo and human in vitro models. Methods: Eighteen New Zealand White rabbits were used. Corneal fibrosis was produced by alkali injury. Twenty-four hours after scar formation, cornea received topically either balanced salt solution (BSS; n = 6), polyethylenimine-conjugated gold nanoparticle (PEI2-GNP)-naked plasmid (n = 6) or PEI2-GNP plasmids expressing BMP7 and HGF genes (n = 6)...
February 1, 2018: Investigative Ophthalmology & Visual Science
Emmanuelle Varlet-Marie, Yvon Sterkers, Marina Perrotte, Patrick Bastien
Toxoplasmosis is generally a benign infection caused by the protozoan parasite Toxoplasma gondii but can have severe consequences in fetuses of mothers infected during pregnancy (congenital toxoplasmosis) and immunocompromised individuals. PCR-based diagnostic tests have become crucial for its diagnosis. However, this molecular diagnosis essentially relies upon laboratory-developed methods and suffers from a lack of standardization, leading to great variation in methods and performance among laboratories. With the need for accreditation of clinical microbiological laboratories, the use of commercial PCR kits has become an attractive alternative; but thorough evaluation of newly commercialized kits by proficient groups is necessary before any recommendation can be made to parasitology laboratories by health authorities or learned societies...
February 22, 2018: International Journal for Parasitology
HyeSoon Song, YouChan Bae, SeokChan Park, HyukMan Kwon, HeeSoo Lee, SeongJoon Joh
Loop-mediated isothermal amplification (LAMP) methods to detect chicken infectious anemia virus (CIAV), reticuloendotheliosis virus (REV), and Marek's disease virus (MDV), and a reverse transcription (RT)-LAMP assay to detect infectious bursal disease virus (IBDV), were developed. The CIAV-LAMP, REV-LAMP, MDV-LAMP, and IBDV-RT-LAMP methods were performed using four sets of six primers targeting the VP1 gene of CIAV, the gp90 gene of REV, the Meq gene of MDV, and the VP2 gene of IBDV. The results (a change in color) were observed visually...
February 21, 2018: Journal of Virological Methods
Kristen L Burkhalter, Keenan Wiggins, Nathan Burkett-Cadena, Barry W Alto
Commercially available assays utilizing antigen or nucleic acid detection chemistries provide options for mosquito control districts to screen their mosquito populations for arboviruses and make timely operational decisions regarding vector control. These assays may be utilized even more advantageously when combined with honey-soaked nucleic acid preservation substrate ('honey card') testing by reducing or replacing the time- and labor-intensive efforts of identifying and processing mosquito pools. We tested artificially inoculated honey cards and cards fed upon individually by West Nile virus (WNV) and Zika virus (ZIKV)-infected mosquitoes with three assays to compare detection rates and the limit of detection for each platform with respect to virus detection of a single infected mosquito and quantify the time interval of virus preservation on the cards...
February 15, 2018: Journal of Medical Entomology
Nichola Eliza Davies Calvani, Sarah Deanna George, Peter Andrew Windsor, Russell David Bush, Jan Šlapeta
Fasciolosis due to infection with Fasciola hepatica, Fasciola gigantica or their hybrids is a significant global cause of livestock production loss. Infection is commonly diagnosed by a labour-intensive sedimentation and faecal egg count (FEC), which has limited throughput and is only applicable after completion of the 8-12 week pre-patent period (PPP). A commercially-available ELISA for the detection of coprological antigen (coproELISA) enables detection prior to the completion of the PPP and is suitable for diagnosis of larger sample sizes, although the sensitivity reported under experimental infection settings can be difficult to replicate in the field, particularly in cattle...
February 15, 2018: Veterinary Parasitology
Owen Higgins, Eoin Clancy, Martin Cormican, Teck Wee Boo, Robert Cunney, Terry J Smith
Bacterial meningitis infection is a leading global health concern for which rapid and accurate diagnosis is essential to reduce associated morbidity and mortality. Loop-meditated isothermal amplification (LAMP) offers an effective low-cost diagnostic approach; however, multiplex LAMP is difficult to achieve, limiting its application. We have developed novel real-time multiplex LAMP technology, TEC-LAMP, using Tth endonuclease IV and a unique LAMP primer/probe. This study evaluates the analytical specificity, limit of detection (LOD) and clinical application of an internally controlled multiplex TEC-LAMP assay for detection of leading bacterial meningitis pathogens: Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae...
February 9, 2018: International Journal of Molecular Sciences
Mitchell G Rohatensky, Devon M Livingstone, Paul Mintchev, Heather K Barnes, Steven C Nakoneshny, Douglas J Demetrick, Joseph C Dort, Guido van Marle
BACKGROUND: Oropharyngeal Squamous Cell Carcinoma (OPSCC) is increasing in incidence despite a decline in traditional risk factors. Human Papilloma Virus (HPV), specifically subtypes 16, 18, 31 and 35, has been implicated as the high-risk etiologic agent. HPV positive cancers have a significantly better prognosis than HPV negative cancers of comparable stage, and may benefit from different treatment regimens. Currently, HPV related carcinogenesis is established indirectly through Immunohistochemistry (IHC) staining for p16, a tumour suppressor gene, or polymerase chain reaction (PCR) that directly tests for HPV DNA in biopsied tissue...
February 8, 2018: BMC Cancer
Joanna Brydak-Godowska, Monika Turczyńska, Mariusz Przybyś, Lidia B Brydak, Dariusz Kęcik
PURPOSE: To describe a case series of ocular complications associated with upper respiratory tract infections. METHODS: Four patients aged 21-61 years (three females, one male) had confirmed ocular complications connected with a general upper respiratory tract infection with myalgia and fever. Ophthalmological examination, including a visual acuity test, a slit-lamp exam, intraocular pressure measurements, fluorescein and indocyanine green angiography, optical coherence tomography (OCT), and diagnostic tests for influenza were performed in the patients (RT-PCR, HAI)...
February 8, 2018: Ocular Immunology and Inflammation
Masatsugu Kimura, Isao Teramoto, Chim W Chan, Zulkarnain Md Idris, James Kongere, Wataru Kagaya, Fumihiko Kawamoto, Ryoko Asada, Rie Isozumi, Akira Kaneko
BACKGROUND: Rapid diagnosis of malaria using acridine orange (AO) staining and a light microscope with a halogen lamp and interference filter was deployed in some malaria-endemic countries. However, it has not been widely adopted because: (1) the lamp was weak as an excitation light and the set-up did not work well under unstable power supply; and, (2) the staining of samples was frequently inconsistent. METHODS: The halogen lamp was replaced by a low-cost, blue light-emitting diode (LED) lamp...
February 7, 2018: Malaria Journal
Jinfeng Sun, Bin Wang, Youjuan Hao, Xueli Yang
This study investigated the effects of calcium dobesilate on Nrf2, Keap1 and HO-1 in the lenses of D-galactose-induced cataracts in rats. Thirty Sprague-Dawley rats were randomly divided into three groups: a blank control group, a model control group and a model administration group. A normal diet was given to the rats in the blank control group and the rats with D-galactose-induced cataracts of the model control group. Calcium dobesilate was also given to the rats with D-galactose-induced cataracts of the model administration group...
January 2018: Experimental and Therapeutic Medicine
Bruce H Noden, Jaclyn Martin, Yisel Carrillo, Justin L Talley, Francisco M Ochoa-Corona
BACKGROUND: The importance of tick and flea-borne rickettsia infections is increasingly recognized worldwide. While increased focus has shifted in recent years to the development of point-of-care diagnostics for various vector-borne diseases in humans and animals, little research effort has been devoted to their integration into vector surveillance and control programs, particularly in resource-challenged countries. One technology which may be helpful for large scale vector surveillance initiatives is loop-mediated isothermal amplification (LAMP)...
2018: PloS One
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