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Multiplex PCR

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https://www.readbyqxmd.com/read/28527284/-new-multiplex-pcr-for-species-specific-diagnosis-of-human-candidiasis
#1
Liliana Torcoroma García, Liany Johanna Luna, Tania Katherine Velasco, Beatriz Elena Guerra
INTRODUCTION: Candidiases is a group of opportunistic infections caused by yeasts belonging to the genus Candida. Candida albicans is the most prevalent species in both superficial and deep infections, however, the clinical importance of non-albicans Candida has increased during the last decade, driving an urgent need for diagnostic tests that allow for species-level resolution and selection of the optimum therapeutic approach. OBJECTIVE: To design and to optimize a new multiplex PCR assay for the simultaneous identification of the five most relevant species of Candida involved in human candidiasis etiology...
June 1, 2017: Biomédica: Revista del Instituto Nacional de Salud
https://www.readbyqxmd.com/read/28526032/multiplex-dnase-free-one-step-reverse-transcription-pcr-for-plasmodium-18s-rrna-and-spliced-gametocyte-specific-mrnas
#2
Amelia E Hanron, Zachary P Billman, Annette M Seilie, Tayla M Olsen, Matthew Fishbaugher, Ming Chang, Thomas Rueckle, Nicole Andenmatten, Bryan Greenhouse, Emmanuel Arinaitwe, John Rek, Smita Das, Gonzalo J Domingo, Kelly Shipman, Stefan H Kappe, James G Kublin, Sean C Murphy
BACKGROUND: Plasmodium gametocytes are sexual stages transmitted to female Anopheles mosquitoes. While Plasmodium parasites can be differentiated microscopically on Giemsa-stained blood smears, molecular methods are increasingly used because of their increased sensitivity. Molecular detection of gametocytes requires methods that discriminate between asexual and sexual stage parasites. Commonly tested gametocyte-specific mRNAs are pfs25 and pfs230 detected by reverse transcription polymerase chain reaction (RT-PCR)...
May 19, 2017: Malaria Journal
https://www.readbyqxmd.com/read/28525762/minimal-residual-disease-monitoring-of-acute-myeloid-leukemia-by-massively-multiplex-digital-pcr-in-patients-with-npm1-mutations
#3
Nuria Mencia-Trinchant, Yang Hu, Maria Antonina Alas, Fatima Ali, Bas J Wouters, Sangmin Lee, Ellen K Ritchie, Pinkal Desai, Monica L Guzman, Gail J Roboz, Duane C Hassane
The presence of minimal residual disease (MRD) is widely recognized as a powerful predictor of therapeutic outcome in acute myeloid leukemia (AML), but methods of measurement and quantification of MRD in AML are not yet standardized in clinical practice. There is an urgent, unmet need for robust and sensitive assays that can be readily adopted as real-time tools for disease monitoring. NPM1 frameshift mutations are an established MRD marker present in half of patients with cytogenetically normal AML. However, detection is complicated by the existence of hundreds of potential frameshift insertions, clonal heterogeneity, and absence of sequence information when the NPM1 mutation is identified using capillary electrophoresis...
May 16, 2017: Journal of Molecular Diagnostics: JMD
https://www.readbyqxmd.com/read/28525287/occurrence-of-enterococcus-faecalis-and-enterococcus-faecium-in-various-clinical-infections-detection-of-their-drug-resistance-and-virulence-determinants
#4
Ali Jahansepas, Mohammad Aghazadeh, Mohammad Ahangarzadeh Rezaee, Alka Hasani, Yaeghob Sharifi, Toofan Aghazadeh, Jalal Mardaneh
The aim of this study was to characterize virulence determinants and antibiotic resistance profiles in enterococci obtained from various clinical sources in the northwest of Iran. A total of 160 enterococcal clinical isolates from various wards of University Teaching Hospitals were collected and specified by biochemical test, from September 2014 to July 2015. Identification of enterococci was confirmed by multiplex PCR in the genus and species level. Antibiotic resistance properties and virulence determinants were examined by phenotypic and molecular methods...
May 19, 2017: Microbial Drug Resistance: MDR: Mechanisms, Epidemiology, and Disease
https://www.readbyqxmd.com/read/28521001/etiology-of-travellers-diarrhea
#5
Z D Jiang, H L DuPont
Eleven published studies of the etiology of travellers' diarrhea (TD) were reviewed define the etiology of TD and to exam newly developed technology such as Real-Time multiplex polymerase chain reaction (PCR) to identify multiple pathogens in one assay to define the cause of TD. Using PCR methods bacterial pathogens were found in 72% of patients acquiring diarrhea in Latin America and in 80% in travellers with illness acquired in Southeast Asia). In these studies, enterotoxigenic Escherichia coli as the predominant pathogen (42% in Latin America and 28% in Southeast Asia)...
April 1, 2017: Journal of Travel Medicine
https://www.readbyqxmd.com/read/28520995/new-molecular-diagnostic-tools-in-traveller-s-diarrhea
#6
Jordi Vila
Traveller's diarrhea can be caused by bacteria, protozoa, helminths and viruses. Globally, the most common causes of traveller's diarrhea are two pathotypes of Escherichia coli (enterotoxigenic and enteroaggregative) and Shigella, although there are significant variations according to the geographic area visited. While traveller's diarrhea is usually a mild, self-limiting disease, half of the travellers with traveller's diarrhea have some limitation in their activities during the journey and up to 10% present persistent diarrhea or other complications, making microbiological diagnosis necessary...
April 1, 2017: Journal of Travel Medicine
https://www.readbyqxmd.com/read/28516828/impact-on-the-medical-decision-making-process-of-multiplex-pcr-assay-for-respiratory-pathogens
#7
A Guillon, S Aymeric, C Gaudy-Graffin, J Sonke, M P Deblonde, J Capsec, P Diot, D Garot, A Goudeau, L Grammatico-Guillon
The objective of this study was to determine how clinicians make use of the modern multiplex PCR assays (MPAs) to manage patients hospitalized for community-acquired pneumonia (CAP). We studied the use of MPAs in 1648 patients hospitalized for CAP over a 3-year period at the moment of the setup of the new PCR assay. We observed that the use of MPAs for the identification of multiple respiratory pathogens marks a radical change in the investigation of CAP etiology. Surprisingly, the contribution of MPAs to the medical decision-making process varies drastically according to the units of care...
May 18, 2017: Epidemiology and Infection
https://www.readbyqxmd.com/read/28516200/respiratory-viral-infections-are-underdiagnosed-in-patients-with-suspected-sepsis
#8
L R Ljungström, G Jacobsson, B E B Claesson, R Andersson, H Enroth
The study aim was to investigate the prevalence and clinical relevance of viral findings by multiplex PCR from the nasopharynx of clinically septic patients during a winter season. During 11 weeks of the influenza epidemic period in January-March 2012, consecutive adult patients suspected to be septic (n = 432) were analyzed with cultures from blood and nasopharynx plus multiplex PCR for respiratory viruses on the nasopharyngeal specimen. The results were compared with those from microbiology analyses ordered as part of standard care...
May 17, 2017: European Journal of Clinical Microbiology & Infectious Diseases
https://www.readbyqxmd.com/read/28515641/molecular-typing-and-antimicrobial-susceptibility-of-staphylococcus-aureus-strains-isolated-from-raw-milk-cheese-minced-meat-and-chicken-meat-samples
#9
Hayriye Yeşim Can, Mehmet Elmalı, Alper Karagöz
The objectives of this study were: i) to detect the presence of Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) in raw milk, cheese, beef minced meat, and chicken meat samples; ii) to evaluate the antimicrobial susceptibility of the isolates; and iii) to determine clonal relation among the isolates by using pulsed-field gel electrophoresis (PFGE) method. Therefore, a total of 160 food samples were randomly collected between August 2014 and May 2015 in Hatay province, located in the southern Turkey...
2017: Korean Journal for Food Science of Animal Resources
https://www.readbyqxmd.com/read/28515454/a-quantitative-shrna-screen-identifies-atp1a1-as-a-gene-that-regulates-cytotoxicity-by-aurilide-b
#10
Shohei Takase, Rumi Kurokawa, Daisuke Arai, Kind Kanemoto Kanto, Tatsufumi Okino, Yoichi Nakao, Tetsuo Kushiro, Minoru Yoshida, Ken Matsumoto
Genome-wide RNA interference (RNAi) with pooled and barcoded short-hairpin RNA (shRNA) libraries provides a powerful tool for identifying cellular components that are relevant to the modes/mechanisms of action (MoA) of bioactive compounds. shRNAs that affect cellular sensitivity to a given compound can be identified by deep sequencing of shRNA-specific barcodes. We used multiplex barcode sequencing technology by adding sample-specific index tags to PCR primers during sequence library preparation, enabling parallel analysis of multiple samples...
May 17, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28513665/a-fully-integrated-microchip-system-for-automated-forensic-short-tandem-repeat-analysis
#11
Junping Han, Wupeng Gan, Bin Zhuang, Jing Sun, Lei Zhao, Jian Ye, Yao Liu, Cai-Xia Li, Peng Liu
We have successfully developed an integrated microsystem that combines two plastic microchips for DNA extraction and PCR amplification with a glass capillary array electrophoresis chip together in a compact control and detection instrument for automated forensic short tandem repeat (STR) analysis. DNA extraction followed by an "in situ PCR" was conducted in a single reaction chamber of the microchip based on a filter paper-based extraction methodology. PCR products were then mixed with sizing standards by an injection electrode and injected into the electrophoresis chip for four-color confocal fluorescence detection...
May 17, 2017: Analyst
https://www.readbyqxmd.com/read/28512861/rapid-diagnosis-of-sepsis-with-taqman-based-multiplex-real-time-pcr
#12
Chang-Feng Liu, Xin-Ping Shi, Yun Chen, Ye Jin, Bing Zhang
BACKGROUND: The survival rate of septic patients mainly depends on a rapid and reliable diagnosis. A rapid, broad range, specific and sensitive quantitative diagnostic test is the urgent need. Thus, we developed a TaqMan-Based Multiplex real-time PCR assays to identify bloodstream pathogens within a few hours. METHODS: Primers and TaqMan probes were designed to be complementary to conserved regions in the 16S rDNA gene of different kinds of bacteria. To evaluate accurately, sensitively, and specifically, the known bacteria samples (Standard strains, whole blood samples) are determined by TaqMan-Based Multiplex real-time PCR...
May 17, 2017: Journal of Clinical Laboratory Analysis
https://www.readbyqxmd.com/read/28511895/diagnostic-performance-of-anyplex-ii-mtb-mdr-xdr-for-detection-of-resistance-to-first-and-second-line-drugs-in-mycobacterium-tuberculosis
#13
F Pérez-García, M J Ruiz-Serrano, P López Roa, Fermín Acosta, Laura Pérez-Lago, D García-De-Viedma, E Bouza
PURPOSE: Genotypic methods have considerably improved the diagnosis of multidrug-resistant (MDR) tuberculosis. One of these tests is Anyplex II MTB/MDR/XDR (Anyplex). Our aim was to evaluate the diagnostic performance of this multiplex PCR. METHODS: We conducted our study on 47 MDR tuberculosis and 14 pan-susceptible strains. We evaluated the ability of Anyplex to detect resistance mutations in rpoB (rifampin [RIF]), katG and inhA (isoniazid [INH]), gyrA (fluoroquinolones [FLQ]), and rrs and eis (aminoglycosides [AMG])...
May 13, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28508913/digitally-encoded-silica-microparticles-for-multiplexed-nucleic-acid-detection
#14
Weiwei Xu, Chao Chen, Xiaodong Ma, Lihua Yuan, Shenquan Liu, Kexiao Zheng, Jiong Li
A robust high throughput suspension array was proposed to analyze biomolecules. To exploit its capacity for nucleic acid detection, we designed a multiplex nested asymmetric PCR (MNAS PCR) that can produce single stranded DNA (ssDNA) efficiently without complicated optimization. Multiplexed HPV genotyping was demonstrated with high selectivity, high sensitivity and rapid hybridization within 20 minutes.
May 16, 2017: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/28507417/molecular-detection-of-taenia-spp-in-dogs-feces-in-zanjan-province-northwest-of-iran
#15
Mohammad Hasan Kohansal, Abbasali Nourian, Ali Haniloo, Asghar Fazaeli
AIM: Echinococcus and Taenia spp. are important but neglected zoonotic helminths of dogs. Dogs as the most relevant definitive hosts harbor several species of Taenia and Echinococcus simultaneously in their gastrointestinal lumen which are morphologically indistinguishable. In this study, we used a multiplex polymerase chain reaction (PCR) method to identify Taeniid infections which seem to be highly distributed in the study region. MATERIALS AND METHODS: A total of 450 dog fecal samples were collected from eight different areas of Zanjan province, northwest of Iran, and examined using a flotation method followed by multiplex PCR for detection and identification of parasites' eggs...
April 2017: Veterinary World
https://www.readbyqxmd.com/read/28506887/molecular-investigation-of-virulence-factors-of-brucella-melitensis-and-brucella-abortus-strains-isolated-from-clinical-and-non-clinical-samples
#16
Reza Mirnejad, Faramarz Masjedian Jazi, Shayan Mostafaei, Mansour Sedighi
Brucella is zoonotic pathogen that induces abortion and sterility in domestic mammals and chronic infections in humans called Malta fever. It is a facultative intracellular potential pathogen with high infectivity. The virulence of Brucella is dependent upon its potential virulence factors such as enzymes and cell envelope associated virulence genes. The aim of this study was to investigate the Brucella virulence factors among strains isolated from humans and animals in different parts of Iran. Seventy eight strains of Brucella species isolated from suspected human and animal cases from several provinces of Iran during 2015-2016 and identified by phenotypic and molecular methods...
May 12, 2017: Microbial Pathogenesis
https://www.readbyqxmd.com/read/28506737/detection-of-genital-chlamydial-and-gonococcal-infection-using-urine-samples-a-community-based-study-from-india
#17
Anjana Krishnan, Sasidharanpillai Sabeena, Parvati V Bhat, Veena Kamath, Maity Hindol, Vahid Rajabali Zadeh, Govindakarnavar Arunkumar
Sexually transmitted infections (STI) have a major impact on the reproductive health of women. Among the different etiological agents of STIs, Chlamydia trachomatis and Neisseria gonorrhoeae are the main bacterial pathogens that cause sexually transmitted infections in women. The aim of the study was to estimate the prevalence of genital chlamydial and gonococcal infection among women in the age group of 18-65 years from a community-based setting. A community-based cross-sectional study was performed using the archived urine samples (n=811) of women in the age group of 18-65 years for C...
May 12, 2017: Journal of Infection and Public Health
https://www.readbyqxmd.com/read/28506262/identification-and-quantification-of-virulence-factors-of-enterotoxigenic-escherichia-coli-by-high-resolution-melting-curve-quantitative-pcr
#18
Weilan Wang, Ruurd T Zijlstra, Michael G Gänzle
BACKGROUND: Diagnosis of enterotoxigenic E. coli (ETEC) associated diarrhea is complicated by the diversity of E.coli virulence factors. This study developed a multiplex quantitative PCR assay based on high-resolution melting curves analysis (HRM-qPCR) to identify and quantify genes encoding five ETEC fimbriae related to diarrhea in swine, i.e. K99, F41, F18, F6 and K88. METHODS: Five fimbriae expressed by ETEC were amplified in multiple HRM-qPCR reactions to allow simultaneous identification and quantification of five target genes...
May 15, 2017: BMC Microbiology
https://www.readbyqxmd.com/read/28506035/simultaneous-molecular-detection-of-cryptosporidium-and-cyclospora-from-raw-vegetables-in-korea
#19
Seobo Sim, Jua Won, Jae-Whan Kim, Kyungjin Kim, Woo-Yoon Park, Jae-Ran Yu
Cryptosporidium and Cyclospora are well-known coccidian protozoa that can cause waterborne and foodborne diarrheal illnesses. There have been a few reports regarding contamination in different vegetables with Cryptosporidium, but no data are available regarding the sources of Cyclospora infections in Korea. In the present study, we collected 6 kinds of vegetables (perilla leaves, winter-grown cabbages, chives, sprouts, blueberries, and cherry tomatoes) from July 2014 to June 2015, and investigated contamination by these 2 protozoa using multiplex quantitative real-time PCR...
April 2017: Korean Journal of Parasitology
https://www.readbyqxmd.com/read/28505584/development-of-a-gold-nanoparticle-based-universal-oligonucleotide-microarray-for-multiplex-and-low-cost-detection-of-foodborne-pathogens
#20
Xiaoqiang Wang, Sisi Ying, Xiaoguang Wei, Jun Yuan
Bacterial foodborne diseases remain major threats to food safety and public health, especially in developing countries. In this study a novel assay, combining gold nanoparticle (GNP)-based multiplex oligonucleotide ligation-PCR and universal oligonucleotide microarray technology, was developed for inexpensive, specific, sensitive, and multiplex detection of eight common foodborne pathogens, including Shigella spp., Campylobacter jejuni, Bacillus cereus, Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica, Staphylococcus aureus, and Vibrio parahaemolyticus...
May 9, 2017: International Journal of Food Microbiology
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