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https://www.readbyqxmd.com/read/29119076/are-there-benefits-in-early-diagnosis-of-prosthetic-joint-infection-with-multiplex-polymerase-chain-reaction
#1
Christian Lausmann, Akos Zahar, Mustafa Citak, Julian Brañes, Stefan Schmidl, Lars Frommelt, Thorsten Gehrke, Matthias Gebauer
Purpose Identification of bacteria and susceptibility are fundamental in periprosthetic joint infection (PJI). Especially in the case of systemic inflammatory response syndrome (SIRS) rapid detection of pathogens is essential for proper therapy. Bacterial cultures are time consuming. The polymerase chain reaction (PCR) is a non-culture molecular method and is able to rapidly identify pathogens and their resistance genes. Multiplex PCR (mPCR) can amplify several different DNA sequences simultaneously. The aim of this study was to show the value of mPCR for early diagnosis of PJI...
2017: Journal of bone and joint infection
https://www.readbyqxmd.com/read/29058256/simultaneous-detection-and-characterization-of-toxigenic-clostridium-difficile-directly-from-clinical-stool-specimens
#2
Hanjiang Lai, Chen Huang, Jian Cai, Julian Ye, Jun She, Yi Zheng, Liqian Wang, Yelin Wei, Weijia Fang, Xianjun Wang, Yi-Wei Tang, Yun Luo, Dazhi Jin
We employed a multiplex polymerase chain reaction (PCR) coupled with capillary electrophoresis (mPCR-CE) targeting six Clostridium difficile genes, including tpi, tcdA, tcdB, cdtA, cdtB, and a deletion in tcdC for simultaneous detection and characterization of toxigenic C. difficile directly from fecal specimens. The mPCR-CE had a limit of detection of 10 colony-forming units per reaction with no cross-reactions with other related bacterial genes. Clinical validation was performed on 354 consecutively collected stool specimens from patients with suspected C...
October 17, 2017: Frontiers of Medicine
https://www.readbyqxmd.com/read/29022552/multiplex-pcr-for-identification-of-six-clinically-relevant-streptococci
#3
Rujirat Hatrongjit, Yukihiro Akeda, Shigeyuki Hamada, Marcelo Gottschalk, Anusak Kerdsin
PURPOSE: The aim of this study was to develop a multiplex PCR (mPCR) for simultaneous detection (single reaction) of six clinically relevant streptococcal species: Streptococcus pneumoniae, Streptococcus suis, Streptococcus gallolyticus subsp. gallolyticus, S. gallolyticus subsp. pasteurianus, Streptococcus intermedius and Streptococcus anginosus/constellatus. METHODS: mPCR with primers specific for S. pneumoniae (lytA), S. suis (recN), S. gallolyticus subsp. gallolyticus (tanB), S...
October 12, 2017: Journal of Medical Microbiology
https://www.readbyqxmd.com/read/28963596/low-occurrence-of-pathogenic-yersinia-enterocolitica-in-pig-tonsils-at-slaughter-in-southern-brazil
#4
Paula Wildemann, Danielle Gava, Ricardo Antônio Pilegi Sfaciotte, Fernanda Danielle Melo, Sandra Maria Ferraz, Ubirajara Maciel da Costa, Eliana Knackfuss Vaz
Yersinia enterocolitica is a foodborne pathogen and pigs are the main reservoir of it in their tonsils. As Brazil is a large producer and exporter of pork meat and information regarding this pathogen is still quite scarce, this study aimed at evaluating the direct detection of Y. enterocolitica followed by pathogenic Y. enterocolitica (PYE) determination in tonsils of slaughtered pigs. For this purpose, 400 pig tonsils were collected from 15 farms in four federally certified slaughterhouses in Southern Brazil...
September 29, 2017: Tropical Animal Health and Production
https://www.readbyqxmd.com/read/28960631/enhanced-single-tube-multiplex-pcr-assay-for-detection-and-identification-of-six-arcobacter-species
#5
I U H Khan, M Cloutier, M Libby, D R Lapen, G Wilkes, E Topp
AIM: A single-tube multiplex PCR (mPCR) assay was developed for rapid, sensitive and simultaneous detection and identification of six Arcobacter species including two new species, A. lanthieri and A. faecis, along with A. butzleri, A. cibarius, A. cryaerophilus and A. skirrowii on the basis of differences in the lengths of their PCR products. Previously designed monoplex, mPCR and RFLP assays do not detect or differentiate A. faecis and A. lanthieri from other closely related known Arcobacter spp...
September 27, 2017: Journal of Applied Microbiology
https://www.readbyqxmd.com/read/28960122/intraocular-detection-of-herpesviruses-by-xtag-liquid-chip-technology-in-patients-with-acute-retinal-necrosis
#6
Chan Zhao, Jie Yi, Fangtian Dong, Rongping Dai, Hanyi Min, Lin Zheng, Youxin Chen, Junjie Ye, Yupei Zhao, Naili Wang, Yingchun Xu, Meifen Zhang
PURPOSE: To evaluate the performance of the xTAG liquid chip technology (xTAG-LCT) for etiological diagnosis of acute retinal necrosis (ARN). METHODS: Fifteen vitreous and 3 aqueous samples from 18 ARN patients were analyzed by xTAG-LCT and multiplex PCR (mPCR)/quantitative PCR (qPCR). RESULTS: xTAG-LCT revealed positive results in 17 of the 18 samples: 10 for Varicella Zoster Virus (VZV) alone; 5 for VZV and Epstein-Barr virus (EBV); 1 for herpes simplex viruses type 1 (HSV-1) and EBV; 1 for VZV, HSV-1 and EBV...
September 29, 2017: Ocular Immunology and Inflammation
https://www.readbyqxmd.com/read/28950041/development-of-a-multiplex-pcr-assay-for-detection-of-pseudomonas-fluorescens-with-biofilm-formation-ability
#7
Yu Xu, Wanyi Chen, Chunping You, Zhenmin Liu
Under the cold storage and processing conditions of raw milk, the psychrotrophic Pseudomonas fluorescens is usually found as predominant bacteria causing its spoilage. In this study, a multiplex PCR assay was developed for rapid and selective detection of P. fluorescens with biofilm formation ability. The target sequences were 2 genes (adnA and fliC) related to biofilm formation and flagella biosynthesis of P. fluorescens. The specificity of the mPCR assay was evaluated with 7 reference strains, isolated from raw milk, belonging to P...
October 2017: Journal of Food Science
https://www.readbyqxmd.com/read/28919163/multiplex-pcr-and-ngs-based-identification-of-mrna-splicing-variants-analysis-of-brca1-splicing-pattern-as-a-model
#8
Jan Hojny, Petra Zemankova, Filip Lhota, Jan Sevcik, Viktor Stranecky, Hana Hartmannova, Katerina Hodanova, Ondrej Mestak, David Pavlista, Marketa Janatova, Jana Soukupova, Michal Vocka, Zdenek Kleibl, Petra Kleiblova
Alternative pre-mRNA splicing increases transcriptome plasticity by forming naturally-occurring alternative splicing variants (ASVs). Alterations of splicing processes, caused by DNA mutations, result in aberrant splicing and the formation of aberrant mRNA isoforms. Analyses of hereditary cancer predisposition genes reveal many DNA variants with unknown clinical significance (VUS) that potentially affect pre-mRNA splicing. Therefore, a comprehensive description of ASVs is an essential prerequisite for the interpretation of germline VUS in high-risk individuals...
December 30, 2017: Gene
https://www.readbyqxmd.com/read/28906580/comparison-of-one-commercial-and-two-in-house-taqman-multiplex-real-time-pcr-assays-for-detection-of-enteropathogenic-enterotoxigenic-and-enteroaggregative-escherichia-coli
#9
COMPARATIVE STUDY
Andreas Hahn, Marc Luetgehetmann, Olfert Landt, Norbert Georg Schwarz, Hagen Frickmann
OBJECTIVE: Enteropathogenic, enterotoxigenic and enteroaggregative Escherichia coli (EPEC, ETEC, EAEC) are among the most frequent causes of diarrhoea during travel or on military deployments. Cost-efficient and reliable real-time multiplex PCR (mPCR) assays are desirable for surveillance or point prevalence studies in remote and resource-limited tropical settings. We compared one commercial PCR kit and two in-house assays without using a gold standard to estimate sensitivity and specificity of each assay...
November 2017: Tropical Medicine & International Health: TM & IH
https://www.readbyqxmd.com/read/28888870/efficient-differentiation-of-corynebacterium-striatum-corynebacterium-amycolatum-and-corynebacterium-xerosis-clinical-isolates-by-multiplex-pcr-using-novel-species-specific-primers
#10
Carolina S Santos, Juliana N Ramos, Veronica V Vieira, Carina S Pinheiro, Roberto Meyer, Neuza M Alcantara-Neves, Rommel T Ramos, Artur Silva, Raphael Hirata, Liza Felicori, Carlos Ruiz de Alegría Puig, Jesús Navas, Vasco Azevedo, Ana L Mattos-Guaraldi, Luis G C Pacheco
A multiplex-PCR (mPCR) assay was designed with species-specific primers which generate amplicons of 226bp, 434bp and 106bp for differentiating the species C. striatum, C. amycolatum, and C. xerosis, respectively. mPCR results were 100% in agreement with identifications achieved by 16S rRNA and rpoB gene sequencing and by VITEK-MS.
November 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28888869/development-of-a-set-of-multiplex-pcrs-for-detection-of-genes-encoding-cell-wall-associated-proteins-in-staphylococcus-pseudintermedius-isolates-from-dogs-humans-and-the-environment
#11
Nathita Phumthanakorn, Pattrarat Chanchaithong, Nuvee Prapasarakul
Staphylococcus pseudintermedius commonly colonizes the skin of dogs, whilst nasal carriage may occur in humans who are in contact with dogs or the environment of veterinary hospitals. Genes encoding cell wall-associated (CWA) proteins have been described in Staphylococcus aureus but knowledge of their occurrence in S. pseudintermedius is still limited. The aim of the study was to develop a method to detect S. pseudintermedius surface protein genes (sps) encoding CWA proteins, and to examine the distribution of the genes in isolates from different sources...
November 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28877158/antibiotic-use-in-hospitalized-children-with-respiratory-viruses-detected-by-multiplex-polymerase-chain-reaction
#12
Pavel Goriacko, Lisa Saiman, Philip Zachariah
BACKGROUND: Multiplex polymerase chain reaction (mPCR) based methods are increasingly used to detect respiratory pathogens in children. While rapid identification of viruses has been shown to reduce antibiotic use, the impact of detecting specific viruses on antibiotic utilization has not been ascertained. This study compared antibiotic utilization among hospitalized children who tested positive for different respiratory viruses at admission. METHODS: A single-center study of hospitalized children under 21 years of age who tested positive at admission for at least one respiratory virus by mPCR from October 1 2012 to October 1 2015 was performed...
September 5, 2017: Pediatric Infectious Disease Journal
https://www.readbyqxmd.com/read/28799826/multiplex-assay-for-identifying-animal-species-found-in-the-tibetan-area-using-the-mitochondrial-12s-rrna-gene
#13
Jung Nam Lee, MingFeng Jiang, YongLi Wen, ShiLin Li, GuoRong Yuan
Southwestern China has an area with unique natural conditions located in alpine regions at altitudes from 2000 to 5000 m; this area is referred to as the Qinghai-Tibetan plateau (QTP). Unique animals, such as yaks (Bos grunniens), are found extensively on the plateau of Southwestern China due to its unique environment. In recent years, the prevalence of fake meat products such as fake jerky has increased in this area. This research was conducted as an attempt to develop a reliable multiplex polymerase chain reaction (mPCR) detection method for identifying nine animal species found in QTP...
August 11, 2017: Animal Biotechnology
https://www.readbyqxmd.com/read/28781945/profiling-of-virulence-associated-factors-in-shigella-species-isolated-from-acute-pediatric-diarrheal-samples-in-tehran-iran
#14
Sajad Yaghoubi, Reza Ranjbar, Mohammad Mehdi Soltan Dallal, Somayeh Yasliani Fard, Mohammad Hasan Shirazi, Mahmood Mahmoudi
OBJECTIVES: The genus Shigella comprises the most infectious and diarrheagenic bacteria causing severe diseases, mostly in children under five years of age. This study aimed to detect nine virulence genes (ipaBCD, VirA, sen, set1A, set1B, ial, ipaH, stx, and sat) in Shigella species (spp.) using multiplex polymerase chain reaction (MPCR) and to determine the relation of Shigella spp. from pediatric diarrheal samples with hospitalization and bloody diarrhea in Tehran, Iran. METHODS: Shigella spp...
June 2017: Osong Public Health and Research Perspectives
https://www.readbyqxmd.com/read/28755939/multiplex-pcr-coupled-with-propidium-monoazide-for-the-detection-of-viable-cronobacter-sakazakii-bacillus-cereus-and-salmonella-spp-in-milk-and-milk-products
#15
Shuang Yu, Leina Yan, Xin Wu, Fan Li, Dong Wang, Hengyi Xu
Cronobacter sakazakii, Bacillus cereus, and Salmonella spp. are common food-borne pathogens. The aim of this study was to develop a sensitive, specific, and rapid method for the simultaneous detection of these 3 pathogens in milk and milk products. Three specific primers were designed based on ompA, invA, and cesB of C. sakazakii, Salmonella spp. and B. cereus, respectively, for use in a multiplex PCR (mPCR). To eliminate false-positive results, cells were pretreated with propidium monoazide (PMA) for the selective elimination of the genomic DNA of dead cells...
July 26, 2017: Journal of Dairy Science
https://www.readbyqxmd.com/read/28755449/population-structure-and-attribution-of-human-clinical-campylobacter-jejuni-isolates-from-central-europe-to-livestock-and-environmental-sources
#16
J Kovac, B Stessl, N Čadež, I Gruntar, M Cimerman, K Stingl, M Lušicky, M Ocepek, M Wagner, S Smole Možina
Campylobacter jejuni is among the most prevalent causes of human bacterial gastroenteritis worldwide. Domesticated animals and, especially, chicken meat are considered to be the main sources of infections. However, the contribution of surface waters and wildlife in C. jejuni transmission to humans is not well understood. We have evaluated the source attribution potential of a six-gene multiplex PCR (mPCR) method coupled with STRUCTURE analysis on a set of 410 C. jejuni strains isolated from environment, livestock, food and humans in central Europe...
July 28, 2017: Zoonoses and Public Health
https://www.readbyqxmd.com/read/28729548/multiplex-polymerase-chain-reaction-for-pathogen-detection-in-donor-recipient-corneal-transplant-tissue-and-donor-storage-solution
#17
Takehiro Hariya, Kazuichi Maruyama, Sunao Sugita, Masayo Takahashi, Shunji Yokokura, Kota Sato, Yasuhiro Tomaru, Norio Shimizu, Toru Nakazawa
Corneal transplantation is a safe, reliable method of restoring visual acuity in patients with corneal disorders. Although it has a very high success rate, rejection can still occur, especially if the site is infected. Therefore, seeking to find better ways to manage infection risk, this study investigated a new technique, based on multiplex polymerase chain reaction (mPCR), to identify pathogens, including viruses, bacteria, and fungi, in corneal transplantation recipient sites, donor corneas and the donor cornea storage solution...
July 20, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28666619/development-of-a-multiplex-pcr-for-identification-of-%C3%AE-hemolytic-streptococci-relevant-to-human-infections-and-serotype-distribution-of-invasive-streptococcus-agalactiae-in-thailand
#18
Anusak Kerdsin, Rujirat Hatrongjit, Shigeyuki Hamada, Yukihiro Akeda, Marcelo Gottschalk
A multiplex polymerase chain reaction (mPCR) was developed for simultaneous detection (single reaction) of genes specific to five frequent clinically relevant β-hemolytic streptococcal species: Streptococcus pyogenes (Spy1258), Streptococcus agalactiae (cfb and cpn60), Streptococcus dysgalactiae subsp. equisimilis (16S-23S intergenic spacer) , S. equi subsp. zooepidemicus (esaA and sorD), and Streptococcus anginosus group (moaC). No cross-reaction was observed with other bacterial species. This test was validated and successfully used with 725 clinical isolates involved in pathological conditions in Thailand and collected between March 2014 and December 2015...
December 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28638437/a-multiplex-pcr-for-rapid-identification-of-brassica-species-in-the-triangle-of-u
#19
Joshua C O Koh, Denise M Barbulescu, Sally Norton, Bob Redden, Phil A Salisbury, Sukhjiwan Kaur, Noel Cogan, Anthony T Slater
BACKGROUND: Within the Brassicaceae, six species from the genus Brassica are widely cultivated throughout the world as oilseed, condiment, fodder or vegetable crops. The genetic relationships among the six Brassica species are described by U's triangle model. Extensive shared traits and diverse morphotypes among Brassica species make identification and classification based on phenotypic data alone challenging and unreliable, especially when dealing with large germplasm collections. Consequently, a major issue for genebank collections is ensuring the correct identification of species...
2017: Plant Methods
https://www.readbyqxmd.com/read/28629316/a-multiplex-primer-design-algorithm-for-target-amplification-of-continuous-genomic-regions
#20
Ahmet Rasit Ozturk, Tolga Can
BACKGROUND: Targeted Next Generation Sequencing (NGS) assays are cost-efficient and reliable alternatives to Sanger sequencing. For sequencing of very large set of genes, the target enrichment approach is suitable. However, for smaller genomic regions, the target amplification method is more efficient than both the target enrichment method and Sanger sequencing. The major difficulty of the target amplification method is the preparation of amplicons, regarding required time, equipment, and labor...
June 19, 2017: BMC Bioinformatics
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