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Caroline H Reuter, Hannah L Palac, Larry K Kociolek, Xiaotian T Zheng, Yusuf Y Chao, Rupal Patel, Sameer J Patel
BACKGROUND: Implementing matrix assisted laser desorption ionization time of flight (MALDI-TOF) and multiplex PCR (mPCR) has been associated with decreased mortality and hospital length of stay in adults, but the impact in pediatrics is less understood. METHODS: This pre-post quasi-experimental study compared antibiotic prescribing for positive blood cultures in patients ≤21 years collected in 2012 (pre-intervention) and in 2015 (after MALDI-TOF/mPCR). Time to effective and optimal antimicrobial therapy were evaluated using Cox proportional hazards regression...
May 10, 2018: Pediatric Infectious Disease Journal
Monika Harms, Volker Schmidt, Tilo Heydel, Jutta Hauptmann, Christine Ahlers, Rene Bergmann, Christoph G Baums
Avian intestinal spirochetosis (AIS), an important but neglected disease in laying hens, is caused by Brachyspira pilosicoli, B. intermedia, and B. alvinipulli. Poultry are also frequently colonized by putatively nonpathogenic species such as B. murdochii and B. innocens. We evaluated the differentiation of Brachyspira species by 3 methods: sequencing of the reduced nicotinamide adenine dinucleotide (NADH) oxidase gene ( nox), matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), and a new multiplex (m)PCR targeting genes such as the tryptophanase A gene ( tnaA) and the p-aminobenzoyl-glutamate hydrolase subunit B gene ( abgB)...
April 1, 2018: Journal of Veterinary Diagnostic Investigation
Xiaoyu Deng, Jiali Zhang, Jiazi Su, Hao Liu, Yanlong Cong, Lei Zhang, Kemeng Zhang, Ning Shi, Rongguang Lu, Xijun Yan
The aim of this study was to establish a multiplex PCR (mPCR) method that can simultaneously detect canine parvovirus (CPV-2), canine coronavirus (CCoV) and canine adenovirus (CAV), thereby eliminating the need to detect these pathogens individually. Based on conserved regions in the genomes of these three viruses, the VP2 gene of CPV-2, the endoribonuclease nsp15 gene of CCoV, and the 52K gene of CAV were selected for primer design. The specificity of the mPCR results showed no amplification of canine distemper virus (CDV), canine parainfluenza virus (CPIV), or pseudorabies virus (PRV), indicating that the method had good specificity...
April 19, 2018: Archives of Virology
Feroze Ganaie, Geetha Nagraj, Vandana Govindan, Reyaz Basha, Mohib Hussain, Nazar Ashraf, Shafique Ahmed, K L Ravikumar
BACKGROUND: The population flow dynamics of Hajj increases the probability of pneumococcal acquisition and amplification among Hajis. This multi-site longitudinal molecular surveillance study was designed to assess the impact and potential variations of pneumococcal carriage in a single cohort of pre and post-Hajj pilgrims from India. METHOD: A total of 3228 pre and post-Hajj, nasopharyngeal and oropharyngeal swabs were collected from 807 pilgrims with an interval of 40 ± 5 days...
April 3, 2018: Travel Medicine and Infectious Disease
Banya Banowary, Van Tuan Dang, Subir Sarker, Joanne H Connolly, Jeremy Chenu, Peter Groves, Shane Raidal, Seyed Ali Ghorashi
Campylobacter infection is a common cause of bacterial gastroenteritis in humans and remains a significant global public health issue. The capability of two multiplex PCR (mPCR)-high-resolution melt (HRM) curve analysis methods (i.e., mPCR1-HRM and mPCR2-HRM) to detect and differentiate 24 poultry isolates and three reference strains of Campylobacter jejuni and Campylobacter coli was investigated. Campylobacter jejuni and C. coli were successfully differentiated in both assays, but the differentiation power of mPCR2-HRM targeting the cadF gene was found superior to that of mPCR1-HRM targeting the gpsA gene or a hypothetical protein gene...
March 2018: Avian Diseases
Muhsin Aydin, Jacqueline Carter-Conger, Ning Gao, David F Gilmore, Steven C Ricke, Soohyoun Ahn
Salmonella is one of major foodborne pathogens and the leading cause of foodborne illness-related hospitalizations and deaths. It is critical to develop a sensitive and rapid detection assay that can identify Salmonella to ensure food safety. In this study, a DNA sensor-based suspension array system of high multiplexing ability was developed to identify eight Salmonella serovars commonly associated with foodborne outbreaks to the serotype level. Each DNA sensor was prepared by activating pre-encoded microspheres with oligonucleotide probes that are targeting virulence genes and serovar-specific regions...
April 2018: Analytical and Bioanalytical Chemistry
M Nour, A Alaidarous
Bacterial meningitis poses enormous healthcare challenges due to a high mortality, morbidity and sequelae. Neisseria (N.) meningitidis, Haemophilus (H.) influenzae, Streptococcus (S.) pneumoniae and S. agalactiae remain among the most prevalent infectious agents that cause bacterial meningitis in children. The objective of this study was the simultaneous detection of these pathogens in suspected cerebrospinal fluid (CSF) by using multiplex polymerase chain reaction (mPCR) and compare PCR results with standard diagnostics currently used in clinical practice...
March 2018: Current Research in Translational Medicine
Jae Ho Sim, Fei Tian, Soo Yeon Jung, Joong-Hyuck Auh, Hyang Sook Chun
Early detection of the zearalenone (ZEA) chemotype of Fusarium species could be a precautionary measure for preventing ZEA contamination in rice. In this study, a multiplex polymerase chain reaction (mPCR) assay for detecting ZEA-producing fungi in rice was established using a set of four primers targeting the ZEA biosynthesis genes PKS3, PKS13, ZEB1, and ZEB2. Two mPCR approaches were used: one that amplified the DNA obtained from Fusarium isolates (conventional method) and another that directly amplified the target DNA from rice samples without time-consuming DNA isolation (direct method)...
March 23, 2018: International Journal of Food Microbiology
Nandagopal Murugan, Jambulingam Malathi, K Lily Therese, Hajib NarahariRao Madhavan
Pseudomonas aeruginosa (P. aeruginosa) is a menacing opportunistic, nosocomial pathogen; become a growing concern as conventional antimicrobial therapy is now futile against it. Multi-drug resistant P. aeruginosa (MDRPA) has distinctive resistance mechanisms such as production of β-lactamases, repression of porin genes and over-expression of efflux pumps. The focus of this study is to standardize and application of multiplex PCR (mPCR) to detect the presence of betalactamase genes encoding blaTem , blaOXA , blaCTX-M-15 , blaVim , blaGes , blaVeb , blaDIM , AmpC and Efflux pump genes encoding Mex A,B-oprM, Mex C,D-oprJ, Mex X,Y-oprN, oprD, nfxB, MexR...
February 2018: Kaohsiung Journal of Medical Sciences
Caijiao Wei, Junliang Zhong, Ting Hu, Xihong Zhao
Escherichia coli O157:H7, Staphylococcus aureus , and Salmonella are food-borne pathogens that cause serious gastrointestinal illness and frequent food safety accidents. This study aimed to develop a practical multiplex polymerase chain reaction (mPCR) technique for the simultaneous detection of these food-borne pathogens in culture broth and artificial food matrix. Pathogen-specific DNA sequences in the rfb E, nuc , and inv A genes were used as targets to design primers for the identification of E. coli O157:H7, S...
January 2018: 3 Biotech
Satsuki Kakiuchi, Masanori Tsuji, Hidekazu Nishimura, Lixing Wang, Mutsuyo Takayama-Ito, Hitomi Kinoshita, Chang-Kweng Lim, Shuichi Taniguchi, Akira Oka, Masashi Mizuguchi, Masayuki Saijo
There have been a few prospective and comprehensive surveillance studies on the respiratory viral infections (RVIs) among patients undergoing hematopoietic stem cell transplantation (HSCT). A 2-year prospective cohort surveillance study of symptomatic and asymptomatic RVIs was performed in hospitalized HSCT patients. Oropharyngeal (OP) swab samples were serially collected each week from 1 week before and up to 100 days after HSCT and were tested for virus isolation with cell culture-based viral isolation (CC-based VI) and a multiplex PCR (MPCR)...
March 22, 2018: Japanese Journal of Infectious Diseases
M Pavlova, V Velev, E Dobreva, G Asseva, I Ivanov, P Petrov, A Mangarov, I Tomova, T Kantardjiev
OBJECTIVE: To develop and optimize a rapid molecular method for diagnosing campylobacteriosis directly from a clinical fecal sample and at the same time for determining the most common causing agents - C. jejuni/coli. MATERIALS AND METHODS: 38 clinical fecal samples from hospitalized patients with diarrheal syndrome were tested using a rapid immunochromatographic test. All positive samples were tested for confirmation by culturing in a microaerophilic atmosphere...
2017: Bratislavské Lekárske Listy
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
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
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...
November 2017: Journal of Medical Microbiology
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...
March 2018: Tropical Animal Health and Production
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
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
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
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
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