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Multiplex PCR

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https://www.readbyqxmd.com/read/28110400/multiplex-pcr-as-a-novel-method-in-the-diagnosis-of-spinal-tuberculosis-a-pilot-study
#1
Kusum Sharma, Rajesh Kumar Meena, Ashish Aggarwal, Rajesh Chhabra
BACKGROUND: Establishment of a reliable and rapid diagnosis is of paramount importance in spinal tuberculosis. The available gadgetry of investigations, such as AFB smear, culture of Mycobacterium tuberculosis, and Uniplex PCR, suffers from a lack of adequate sensitivity and/or a lack of rapidity. Therefore, many times a diagnosis is made either very late in the disease process or sometimes empirical therapy has to be started because a definite diagnosis could not be made. All of these are not ideal situations for a clinician...
January 21, 2017: Acta Neurochirurgica
https://www.readbyqxmd.com/read/28107927/electrochemical-detection-of-magnetically-entrapped-dna-sequences-from-complex-samples-by-multiplexed-enzymatic-labelling-application-to-a-transgenic-food-feed-quantitative-survey
#2
C L Manzanares-Palenzuela, J P Martín-Clemente, M J Lobo-Castañón, B López-Ruiz
Monitoring of genetically modified organisms in food and feed demands molecular techniques that deliver accurate quantitative results. Electrochemical DNA detection has been widely described in this field, yet most reports convey qualitative data and application in processed food and feed samples is limited. Herein, the applicability of an electrochemical multiplex assay for DNA quantification in complex samples is assessed. The method consists of the simultaneous magnetic entrapment via sandwich hybridisation of two DNA sequences (event-specific and taxon-specific) onto the surface of magnetic microparticles, followed by bienzymatic labelling...
March 1, 2017: Talanta
https://www.readbyqxmd.com/read/28107671/recent-sequence-variation-in-probe-binding-site-affected-detection-of-respiratory-syncytial-virus-group-b-by-real-time-rt-pcr
#3
Everlyn Kamau, Charles N Agoti, Clement S Lewa, John Oketch, Betty E Owor, Grieven P Otieno, Anne Bett, Patricia A Cane, D James Nokes
BACKGROUND: Direct immuno-fluorescence test (IFAT) and multiplex real-time RT-PCR have been central to RSV diagnosis in Kilifi, Kenya. Recently, these two methods showed discrepancies with an increasing number of PCR undetectable RSV-B viruses. OBJECTIVES: Establish if mismatches in the primer and probe binding sites could have reduced real-time RT-PCR sensitivity. STUDY DESIGN: Nucleoprotein (N) and glycoprotein (G) genes were sequenced for real-time RT-PCR positive and negative samples...
January 5, 2017: Journal of Clinical Virology: the Official Publication of the Pan American Society for Clinical Virology
https://www.readbyqxmd.com/read/28107507/a-multiplex-rt-pcr-assay-for-detection-and-differentiation-of-avian-origin-canine-h3n2-equine-origin-h3n8-human-origin-h3n2-and-h1n1-2009-canine-influenza-viruses
#4
Chenxi Wang, Qian Wang, Junyi Hu, Honglei Sun, Juan Pu, Jinhua Liu, Yipeng Sun
Virological and serological surveys have documented that H1N1/2009, avian-origin canine H3N2 (cH3N2), seasonal human-origin H3N2 (hH3N2), and equine-origin H3N8 influenza viruses are consistently circulating in dogs. In the present study, a multiplex reverse-transcriptase polymerase chain reaction (mRT-PCR) assay was developed for simultaneous detection and differentiation of these influenza viruses. Four primer sets were designed to target the hemagglutinin genes of H1N1/2009, cH3N2, hH3N2, and H3N8 canine influenza viruses (CIVs)...
2017: PloS One
https://www.readbyqxmd.com/read/28105353/isolated-extramedullary-cutaneous-relapse-despite-concomitant-severe-graft-vs-host-disease-and-tissue-chimerism-analysis-in-a-patient-with-acute-lymphoblastic-leukemia-after-allogeneic-hematopoietic-stem-cell-transplantation-a-case-report
#5
Bulent Kantarcioglu, Huseyin Saffet Bekoz, Yeliz Duvarci Ogret, Asli Cakir, Demet Kivanc, Fatma Savran Oguz, Deniz Sargin
Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is a potentially curative treatment option for patients with acute lymphoblastic leukemia (ALL). The curative potential of allo-HSCT for ALL is, in part, due to the graft-vs.-leukemia (GVL) effect, in addition to the intensive conditioning chemo-radiotherapy. However, relapse remains the major cause of treatment failure following allo-HSCT for ALL. In the allo-HSCT setting, testing for genetic markers of hematopoietic chimerism has become a part of the routine diagnostic program...
December 2016: Molecular and Clinical Oncology
https://www.readbyqxmd.com/read/28100599/rapid-and-specific-detection-of-the-escherichia-coli-sequence-type-648-complex-stc648-within-phylogroup-f
#6
James R Johnson, Brian D Johnston, David M Gordon
The Escherichia coli sequence type 648 complex (STc648) is an emerging lineage within phylogroup F - formerly included within phylogroup D - that is associated with multidrug resistance. Here, we designed and validated a novel multiplex PCR-based assay for STc648 that took advantage of (i) four distinctive single-nucleotide polymorphisms in icd allele 96 and gyrB allele 87, two of the multi-locus sequence typing alleles that define ST648, and (ii) the typical absence within STc648 of uidA, an E. coli-specific gene encoding β-glucuronidase...
January 18, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28100261/serotypes-of-streptococcus-suis-isolated-from-healthy-pigs-in-phayao-province-thailand
#7
P Thongkamkoon, T Kiatyingangsulee, M Gottschalk
BACKGROUND: Streptococcus suis (S. suis) is an important swine and human pathogen. There are 33 serotypes that have been described. Zoonotic cases are very common the Northern part of Thailand, especially in Phayao Province. However, the prevalence of S. suis and, more particularly the different serotypes, in pigs in this region is poorly known and needed to be addressed. THE CONTEXT AND PURPOSE OF THE STUDY: Distribution of S. suis serotypes varies depending on the geographical area...
January 19, 2017: BMC Research Notes
https://www.readbyqxmd.com/read/28100169/medium-throughput-bisulfite-sequencing-for-accurate-detection-of-5-methylcytosine-and-5-hydroxymethylcytosine
#8
Gary G Chen, Jeffrey A Gross, Pierre-Eric Lutz, Kathryn Vaillancourt, Gilles Maussion, Alexandre Bramoulle, Jean-François Théroux, Elena S Gardini, Ulrike Ehlert, Geneviève Bourret, Aurélie Masurel, Pierre Lepage, Naguib Mechawar, Gustavo Turecki, Carl Ernst
BACKGROUND: Epigenetic modifications of DNA, such as 5-methylcytosine and 5-hydroxymethycytosine, play important roles in development and disease. Here, we present a cost-effective and versatile methodology for the analysis of DNA methylation in targeted genomic regions, which comprises multiplexed, PCR-based preparation of bisulfite DNA libraries followed by customized MiSeq sequencing. RESULTS: Using bisulfite and oxidative bisulfite conversion of DNA, we have performed multiplexed targeted sequencing to analyse several kilobases of genomic DNA in up to 478 samples, and achieved high coverage data of 5-methylcytosine and 5-hydroxymethycytosine at single-base resolution...
January 18, 2017: BMC Genomics
https://www.readbyqxmd.com/read/28099766/high-salinity-relaying-to-reduce-vibrio-parahaemolyticus-and-vibrio-vulnificus-in-chesapeake-bay-oysters-crassostrea-virginica
#9
Salina Parveen, Michael Jahncke, Sara Elmahdi, Helen Crocker, John Bowers, Chanelle White, Stephanie Gray, Amanda C Morris, Kathy Brohawn
Cases of Vibrio infections in the United States have tripled from 1996 to 2009 and these infections are most often associated with the consumption of seafood, particularly oysters (Crassostrea virginica). Information is needed on how to reduce numbers of Vibrio parahaemolyticus and Vibrio vulnificus in bi-valve molluscan shellfish (for example, oysters). The purpose of this study was to evaluate the effectiveness of high salinity relaying or treatment in recirculating aquaculture systems (RASs) as methods to reduce the abundance of V...
January 18, 2017: Journal of Food Science
https://www.readbyqxmd.com/read/28099581/minimal-residual-disease-detection-in-tunisian-b-acute-lymphoblastic-leukemia-based-on-immunoglobulin-gene-rearrangements
#10
S Besbes, W S Hamadou, M L Boulland, Y B Youssef, B Achour, H Regaieg, A Khelif, T Fest, Z Soua
IGH gene rearrangement and IGK-Kde gene deletion can be used as molecular markers for the assessment of B lineage acute lymphoblastic leukemia (B-ALL). Minimal residual disease detected based on those markers is currently the most reliable prognosis factor in B-ALL. The aim of this study was to use clonal IGH/IGK-Kde gene rearrangements to confirm B-ALL diagnosis and to evaluate the treatment outcome of Tunisian leukemic patients by monitoring the minimal residual disease (MRD) after induction chemotherapy...
January 16, 2017: Brazilian Journal of Medical and Biological Research, Revista Brasileira de Pesquisas Médicas e Biológicas
https://www.readbyqxmd.com/read/28098526/ancylostoma-ceylanicum-hookworm-in-the-solomon-islands
#11
Richard S Bradbury, Sze Fui Hii, Humpress Harrington, Richard Speare, Rebecca Traub
Although hookworm is highly prevalent in the Solomon Islands, the species involved are unknown. We initiated this study in response to finding Ancylostoma ceylanicum hookworm in a peacekeeper in Australia who had returned from the Solomon Islands. Kato-Katz fecal surveys performed in 2013 and 2014 in 2 village groups in East Malaita, Solomon Islands, identified hookworm-positive samples. These specimens were tested by cytochrome oxidase 1 (cox-1) gene multiplex PCR and sequenced. Of 66 positive specimens, 54 (81...
February 2017: Emerging Infectious Diseases
https://www.readbyqxmd.com/read/28096862/does-nosema-ceranae-wipe-out-nosema-apis-in-turkey
#12
Rahşan Ivgin Tunca, Devrim Oskay, Ayhan Gosterit, Olgay Kaan Tekin
BACKGROUND: The aim of this study was to determine the prevalence of the Nosema ceranae and Nosema apis among apiaries using both spore counts and multiplex PCR and the replacement of N. apis by N. ceranae in some regions of Turkey. METHODS: A hundred honey bee samples were collected from 99 apiaries in 11 different locations in 2011-2012 in Turkey. Nosema infection degree from collected samples was determined using light microscope and molecular detection of Nosema spp...
April 2016: Iranian Journal of Parasitology
https://www.readbyqxmd.com/read/28096860/determination-of-asymptomatic-malaria-among-afghani-and-pakistani-immigrants-and-native-population-in-south-of-kerman-province-iran
#13
Mohammad Bagher Amirshekari, Mehdi Nateghpour, Ahmad Raeisi, Afsaneh Motevalli Haghi, Leila Farivar, Gholamhosein Edrissian
BACKGROUND: This study was proposed to monitor the situation of asymptomatic malaria among the native population and Afghani and Pakistani immigrants in Kahnooj and Ghale-Ganj districts from Kerman Province, Southeastern Iran. METHODS: A number of 180 and 120 individuals from Kahnooj and Ghale-Ganj respectively were registered and considered based on a cross-sectional surveillance method. From 300 registered cases, 200 individuals (66.7%) were selected among Afghani and Pakistani immigrants and the rest (33...
April 2016: Iranian Journal of Parasitology
https://www.readbyqxmd.com/read/28096617/phenotypic-and-genotypic-characterization-of-locally-isolated-salmonella-strains-used-in-preparation-of-salmonella-antigens-in-egypt
#14
Hazem Mohammed Ibrahim, Dalia Ahmed Mohammed Abd El-Moaty, Hanan Ali Ahmed, Mona Ibrahim El-Enbaawy
AIM: This work was conducted to study the phenotypic and genotypic characterization of locally isolated Salmonella strains (Salmonella Pullorum, Salmonella Enteritidis, and Salmonella Typhimurium) from poultry used in the preparation of Salmonella antigens in Egypt. MATERIALS AND METHODS: The phenotypic characterization of Salmonella strains was done using standard microbiological, biochemical, and serological techniques. Molecular identification was done using different sets of primers on different genes using different polymerase chain reaction (PCR) techniques...
December 2016: Veterinary World
https://www.readbyqxmd.com/read/28093224/beta-2-adrenoreceptor-polymorphism-at-position-16-determines-the-clinical-severity-of-chronic-obstructive-pulmonary-disease
#15
Justyna Emeryk-Maksymiuk, Andrzej Emeryk, Paweł Krawczyk, Kamila Wojas-Krawczyk, Janusz Milanowski
The Arg/Arg homozygosity at codon 16 of the beta-2-adrenoreceptor (ADRB2) gene has been thought to predispose asthma patients to a poorer therapeutic response to beta-2-mimetics, or to worse control of the disease. In contrast, the results of the studies analysing the effect of ADRB2 polymorphisms on the response to beta-2-adrenoreceptor agonists in chronic obstructive pulmonary disease (COPD) patients are sparse and inconclusive. The aim of this research was to verify if p.Arg16Gly (c.46A > G) and p.Gly27Glu (c...
January 13, 2017: Pulmonary Pharmacology & Therapeutics
https://www.readbyqxmd.com/read/28091937/ocular-mycobacteriosis-dual-infection-of-m-tuberculosis-complex-with-m-fortuitum-and-m-bovis
#16
Kusum Sharma, Natasha Gautam, Megha Sharma, Mohit Dogra, Priya Bajgai, Basavaraj Tigari, Aman Sharma, Vishali Gupta, Surya Prakash Sharma, Ramandeep Singh
BACKGROUND: We report unfavorable outcome in a patient with subretinal granuloma caused by dual infection of Mycobacterium tuberculosis complex with Mycobacterium fortuitum and Mycobacterium bovis in an immunosuppressed, non-HIV patient. We did a systematic review of literature on dual infection due to M. tuberculosis and M. fortuitum via MEDLINE and PUBMED and could not find any case reported of causing this kind of dual infection in the eye. RESULTS: A 38-year-old Indian male patient presented with decreased vision in the left eye for 3 months, diagnosed as tubercular choroidal granuloma with associated retinal angiomatosis proliferans (RAP) lesion...
December 2017: Journal of Ophthalmic Inflammation and Infection
https://www.readbyqxmd.com/read/28088897/multiplex-pcr-as-a-rapid-and-sensitive-method-for-identification-of-meat-species-in-halal-meat-products
#17
Mahsa Alikord, Javad Keramat, Mahdi Kadivar, Hassan Momtaz, Mohammad Naghi Eshtiaghi, Aziz Homayouni Rad
BACKGROUND: Species identification and authentication in meat products are important subjects for ensuring the health of consumers. The multiplex-PCR amplification and species-specific primer set were used for identification of horse, donkey, pig and other ruminants in raw and processed meat products. METHODS: Oligonucleotid primers were designed for amplification of species-specific mitochondrial DNA sequences of each species and samples were prepared from binary meat mixtures...
January 13, 2017: Recent Patents on Food, Nutrition & Agriculture
https://www.readbyqxmd.com/read/28081347/virulence-factors-biofilm-forming-ability-and-antimicrobial-resistance-of-urinary-escherichia-coli-strains-isolated-from-hospitalized-patients
#18
Barbara Kot, Jolanta Wicha, Agata Grużewska, Małgorzata Piechota, Katarzyna Wolska, Maria Obrębska
BACKGROUND/AIM: Escherichia coli is the most frequent cause of urinary tract infections. We investigated the possible associations between the origin of strains, antimicrobial resistance, the presence of urovirulence factors, and biofilm-forming ability. MATERIALS AND METHODS: Antibiotic susceptibility of E. coli strains was tested by disk diffusion method. Hemagglutination assays were performed for phenotypic characterization of the cell surface. Multiplex PCR was used for detection of virulence genes and for determination of phylogenetic relationships...
December 20, 2016: Turkish Journal of Medical Sciences
https://www.readbyqxmd.com/read/28081126/a-pan-lyssavirus-taqman-real-time-rt-pcr-assay-for-the-detection-of-highly-variable-rabies-virus-and-other-lyssaviruses
#19
Ashutosh Wadhwa, Kimberly Wilkins, Jinxin Gao, Rene Edgar Condori Condori, Crystal M Gigante, Hui Zhao, Xiaoyue Ma, James A Ellison, Lauren Greenberg, Andres Velasco-Villa, Lillian Orciari, Yu Li
Rabies, resulting from infection by Rabies virus (RABV) and related lyssaviruses, is one of the most deadly zoonotic diseases and is responsible for up to 70,000 estimated human deaths worldwide each year. Rapid and accurate laboratory diagnosis of rabies is essential for timely administration of post-exposure prophylaxis in humans and control of the disease in animals. Currently, only the direct fluorescent antibody (DFA) test is recommended for routine rabies diagnosis. Reverse-transcription polymerase chain reaction (RT-PCR) based diagnostic methods have been widely adapted for the diagnosis of other viral pathogens, but there is currently no widely accepted rapid real-time RT-PCR assay for the detection of all lyssaviruses...
January 2017: PLoS Neglected Tropical Diseases
https://www.readbyqxmd.com/read/28078872/molecular-characterization-and-antibiotic-resistance-of-salmonella-serovars-isolated-in-the-apulia-region-of-italy
#20
R Mirgaldi, A Ballini, A M Dionisi, I Luzzi, G Dipalma, F Inchingolo, D De Vito
During the period January 2013-December 2015, 175 cases of human salmonellosis were reported in the Apulia Region of Italy. The aim of this study was to characterize salmonella strains from the standpoints of serovars prevalence, antimicrobial resistance and clonal origin. The serological typing was performed by agglutination against antisera followed by a multiplex polymerase chain reaction (m-PCR). The obtained results were analyzed following the Kauffmann-White scheme. Susceptibility to antimicrobial agents was tested using the disk diffusion method on Muller-Hinton agar plates...
October 2016: Journal of Biological Regulators and Homeostatic Agents
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