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https://www.readbyqxmd.com/read/28729548/multiplex-polymerase-chain-reaction-for-pathogen-detection-in-donor-recipient-corneal-transplant-tissue-and-donor-storage-solution
#1
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
#2
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...
June 27, 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
#3
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
#4
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
https://www.readbyqxmd.com/read/28624018/difference-in-the-craniocaudal-gradient-of-the-maximum-pixel-value-change-rate-between-chronic-obstructive-pulmonary-disease-patients-and-normal-subjects-using-sub-mgy-dynamic-chest-radiography-with-a-flat-panel-detector-system
#5
Yoshitake Yamada, Masako Ueyama, Takehiko Abe, Tetsuro Araki, Takayuki Abe, Mizuki Nishino, Masahiro Jinzaki, Hiroto Hatabu, Shoji Kudoh
OBJECTIVES: To compare the craniocaudal gradients of the maximum pixel value change rate (MPCR) during tidal breathing between chronic obstructive pulmonary disease (COPD) patients and normal subjects using dynamic chest radiography. MATERIALS AND METHODS: This prospective study was approved by the institutional review board and all participants provided written informed consent. Forty-three COPD patients (mean age, 71.6±8.7 years) and 47 normal subjects (non-smoker healthy volunteers) (mean age, 54...
July 2017: European Journal of Radiology
https://www.readbyqxmd.com/read/28615466/a-pathotyping-multiplex-pcr-for-haemophilus-parasuis-a-tool-for-the-prediction-of-virulence
#6
Kate J Howell, Lucy A Weinert, Sarah E Peters, Jinhong Wang, Juan Hernandez-Garcia, Roy R Chaudhuri, Shi-Lu Luan, Øystein Angen, Virginia Aragon, Susanna M Williamson, Paul R Langford, Andrew N Rycroft, Brendan W Wren, Duncan J Maskell, Alexander W Tucker
Haemophilus parasuis is a diverse bacterial species found in the upper respiratory tract of pigs that can also cause Glässer's disease and pneumonia. A previous pan-genome study of H. parasuis identified 48 genes that were associated with clinical disease. Here we describe the development of a generalised linear model (termed a pathotyping model) to predict the potential virulence of isolates of H. parasuis, based on a subset of 10 genes from the pan-genome. A multiplex PCR (mPCR) was constructed based on these genes, the results of which were entered into the pathotyping model to yield a prediction of virulence...
June 14, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28612135/comparative-study-of-genexpert-mtb-rif-assay-and-multiplex-pcr-assay-for-direct-detection-of-mycobacterium-tuberculosis-in-suspected-pulmonary-tuberculosis-patients
#7
Anil Kumar Sah, Bishnu Joshi, Dhruba Kumar Khadka, Birendra Prasad Gupta, Anurag Adhikari, Subodh Kant Singh, Ganesh Rai, Geeta Shrestha Vaidya, Reema Rajbhandari, Basant Pant, Shiba Kumar Rai
Pulmonary tuberculosis (PTB) is one of the major infectious diseases in developing countries. The objective of this study was to compare rapid diagnostics technique, GeneXpert MTB/RIF (GeneXpert) and Multiplex PCR assay (MPCR) targeting IS6110 segment and mpb64 gene for direct detection of Mycobacterium tuberculosis (MTB) in suspected PTB patients. A cross sectional study was carried among 105 sputum samples from suspected PTB patients to evaluate GeneXpert and Multiplex PCR who visited National Tuberculosis Center, Nepal...
June 13, 2017: Current Microbiology
https://www.readbyqxmd.com/read/28584712/prevalence-and-seroepidemiology-of-haemophilus-parasuis-in-sichuan-province-china
#8
Zhenghao Wang, Qin Zhao, Hailin Wei, Xintian Wen, Sanjie Cao, Xiaobo Huang, Rui Wu, Qigui Yan, Yong Huang, Yiping Wen
Haemophilus parasuis, the causative agent of Glässer's disease, has been reported as widespread, but little is known about its epidemiology in the Sichuan province of China. The goal of our research is to reveal the prevalence and distribution of H. parasuis in this area. Sampling and isolation were performed across Sichuan; isolates were processed using serotyping multiplex PCR (serotyping-mPCR) and agar gel diffusion (AGD) for confirmation of serovar identity. This study was carried out from January 2014 to May 2016 and 254 H...
2017: PeerJ
https://www.readbyqxmd.com/read/28574617/confirmed-identification-and-toxin-profiling-of-campylobacter-jejuni-using-a-thermostabilized-multiplex-pcr-formulation
#9
Nitya Ramachandran, Shylaja Ramlal, Harsh Vardhan Batra
Cytolethal distending toxin (CDT) producing Campylobacter jejuni species are one of the leading causes of human gastroenteritis worldwide. The main intent of the study was to develop a multiplex PCR assay for the confirmed identification and toxin profiling of C. jejuni. The genes targeted were rpo B as genus specific, hip O for species; cdt A, cdt B, cdt C encoding respective subunit proteins of CDT with Internal Amplification Control (IAC). To enhance its application as a pre-mixed ready-to-use format, the master mix of developed mPCR was dried by lyophilization and stability was assessed...
July 2017: APMIS: Acta Pathologica, Microbiologica, et Immunologica Scandinavica
https://www.readbyqxmd.com/read/28571452/miniaturized-thermocycler-based-on-thermoelectric-heating-for-diagnosis-of-sexually-transmitted-disease-by-dna-amplification
#10
Hyunjung Lim, Ga Eun Jo, Kyong Soo Kim, Seung Min Back, Hyuk Choi
Sexually transmitted disease (STD) is among the most common infectious diseases; therefore, it is necessary to develop sensitive early diagnostic techniques. As the gold standard, polymerase chain reaction (PCR) has been most widely employed for STD diagnosis; however, PCR requires large and expensive instruments. In this study, miniaturized thermal cycler using Peltier modules was developed for the PCR analysis. In comparison with the conventional PCR instrument, the Peltier-based micro-PCR (P-mPCR) device developed in this study enables one to amplify and successfully distinguish between DNA of different sizes...
May 2017: Review of Scientific Instruments
https://www.readbyqxmd.com/read/28559002/assessment-of-the-automated-multiplex-pcr-unyvero-i60-iti-%C3%A2-cartridge-system-to-diagnose-prosthetic-joint-infection-a-multicenter-study
#11
D Malandain, P Bémer, A G Leroy, J Léger, C Plouzeau, A S Valentin, A Jolivet-Gougeon, D Tandé, G Héry-Arnaud, C Lemarié, M Kempf, L Bret, C Burucoa, S Corvec
OBJECTIVES: Prosthetic joint infections (PJI) are responsible for significant morbidity and mortality and their number continues to rise. Their management remains complex, especially the microbiological diagnosis. Besides "homemade" tests developed by several teams, new molecular biology methods are now available with different analytical performance and usability. METHODS: We studied the performances of one of these tests: ITI(®) multiplex PCR (mPCR) by the Curetis(®) company and compared it either to "optimized" culture or to 16S rRNA PCR...
May 27, 2017: Clinical Microbiology and Infection
https://www.readbyqxmd.com/read/28515602/respiratory-multiplex-polymerase-chain-reaction-an-important-diagnostic-tool-in-immunocompromised-patients
#12
Amarjeet Kaur, Navin Kumar, Sharmila Sengupta, Yatin Mehta
BACKGROUND: Viruses and atypical pathogens can cause significant respiratory illness in immunocompromised patients. Multiplex polymerase chain reaction (MPCR) has improved the diagnostic yield of pathogens, and it is easier to identify the co-infections also. The present study was done to evaluate the performance of MPCR on bronchoalveolar lavage (BAL) samples in immunocompromised patients. METHODS: Atotal of 177 BAL specimens collected over a 19 months period from immunocompromised patients with respiratory illness were analyzed with the MPCR and aerobic culture...
April 2017: Indian Journal of Critical Care Medicine
https://www.readbyqxmd.com/read/28494435/impact-of-respiratory-viruses-in-hospital-acquired-pneumonia-in-the-intensive-care-unit-a-single-center-retrospective-study
#13
Paul Loubet, Guillaume Voiriot, Nadhira Houhou-Fidouh, Mathilde Neuville, Lila Bouadma, Francois-Xavier Lescure, Diane Descamps, Jean-François Timsit, Yazdan Yazdanpanah, Benoit Visseaux
BACKGROUND: Data on the frequency and role of respiratory viruses (RVs) in hospital-acquired pneumonia (HAP) are still scarce. OBJECTIVES: We assessed the proportion of RVs and their impact on the outcome of hospital-acquired pneumonia (HAP) in the intensive care unit (ICU). STUDY DESIGN: Cases of HAP were retrospectively selected among patients who underwent screening for RVs by multiplex PCR (mPCR) in the ICU of a French tertiary care hospital from May 2014 to April 2016...
April 22, 2017: Journal of Clinical Virology: the Official Publication of the Pan American Society for Clinical Virology
https://www.readbyqxmd.com/read/28330075/a-novel-multiplex-pcr-method-for-the-detection-of-virulence-associated-genes-of-escherichia-coli-o157-h7-in-food
#14
Vo Van Giau, Thuy Trang Nguyen, Thi Kim Oanh Nguyen, Thi Thuy Hang Le, Tien Dung Nguyen
Shiga toxin-producing Escherichia coli O157:H7 (E. coli O157:H7) strains are foodborne infectious agents that cause a number of life-threatening diseases, including hemorrhagic colitis (HC) and hemolytic uremic syndrome (HUS). Shiga toxin 1 (stx1), shiga toxin 2 (stx2), or a combination of both are responsible for most clinical symptoms of these diseases. Hence, various diagnostic methods have been developed so far to detect shiga toxins such as cell culture, ELISA, Rapid Latex Agglutination (RPLA) and hybridization, but due to high costs and labor time in addition to low sensitivity, they have not received much attention...
June 2016: 3 Biotech
https://www.readbyqxmd.com/read/28110400/multiplex-pcr-as-a-novel-method-in-the-diagnosis-of-spinal-tuberculosis-a-pilot-study
#15
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...
March 2017: Acta Neurochirurgica
https://www.readbyqxmd.com/read/28005999/characterization-of-chinese-haemophilus-parasuis-isolates-by-traditional-serotyping-and-molecular-serotyping-methods
#16
Lina Ma, Liyan Wang, Yuefeng Chu, Xuerui Li, Yujun Cui, Shengli Chen, Jianhua Zhou, Chunling Li, Zhongxin Lu, Jixing Liu, Yongsheng Liu
Haemophilus parasuis is classified mainly through serotyping, but traditional serotyping always yields non-typable (NT) strains and unreliable results via cross-reactions. Here, we surveyed the serotype prevalence of Chinese H. parasuis isolates using traditional serotyping (gel immuno-diffusion test, GID) and molecular serotyping (multiplex PCR, mPCR). We also investigated why discrepant results between these methods were obtained, and investigated mPCR failure through whole-genome sequencing. Of the 100 isolate tested, 73 (73%) and 93 (93%) were serotyped by the GID test and mPCR, respectively, with a concordance rate of 66% (66/100)...
2016: PloS One
https://www.readbyqxmd.com/read/27923665/nicotine-enhances-gabaergic-inhibition-of-oxytocin-mrna-expressing-neuron-in-the-hypothalamic-paraventricular-nucleus-in-vitro-in-rats
#17
Ke-Min Zhang, Guo-Yan Zhao, Bin-Bin Zhang, Qi Xu, Chun-Ping Chu, Hua Jin, De-Lai Qiu
We recently found that extracellular administration of nicotine indirectly excited hypothalamic paraventricular nucleus (PVN) corticotropin-releasing hormone (CRH) mRNA-expressing neurons. In this study, we studied the effect of nicotine on PVN oxytocin (OT) mRNA-expressing neuron in vitro in rats, by whole-cell patch-clamp recording technique, immunohistochemistry methods and single-cell reverse-transcription multiplex polymerase chain reaction (SC-RT-mPCR) methods Our results showed that 79.3% (73/92) of the 92 PVN putative magnocellular neurons co-expressed GAPDH mRNA and OT mRNA...
December 5, 2016: Neuroscience Letters
https://www.readbyqxmd.com/read/27922429/multiplex-pcr-for-detection-of-helicobacter-pylori-infection-in-gastric-biopsies-with-lower-inflammatory-score
#18
Najmiyatul Fadilah, Alfizah Hanafiah, Hamizah Razlan, Zin Qin Wong, Isa Mohamed Rose, Md Mostafizur Rahman
BACKGROUND: No gold standard has yet been established for the diagnosis of H. pylori infection. A multiplex polymerase chain reaction (mPCR) was developed in this study for rapid, sensitive and specific detection of H. pylori from gastric biopsies. METHODS: H. pylori infections were determined by in-house rapid urease test (iRUT), culture, histology and multiplex PCR. RESULTS: A total of 140 (60.9%) from 230 patients were positive for H. pylori infection...
October 2016: British Journal of Biomedical Science
https://www.readbyqxmd.com/read/27906992/a-dual-filtration-based-multiplex-pcr-method-for-simultaneous-detection-of-viable-escherichia-coli-o157-h7-listeria-monocytogenes-and-staphylococcus-aureus-on-fresh-cut-cantaloupe
#19
Ke Feng, Wenzhong Hu, Aili Jiang, Sarengaowa, Yongping Xu, Yu Zou, Liu Yang, Xin Wang
Fresh-cut cantaloupe is particularly susceptible to contamination with pathogenic bacteria, such as Escherichia coli O157:H7, Listeria monocytogenes, and Staphylococcus aureus. Therefore, development of rapid, yet accurate detection techniques is necessary to ensure food safety. In this study, a multiplex PCR system and propidium monoazide (PMA) concentration were optimized to detect all viable pathogens in a single tube. A dual filtration system utilized a filtration membrane with different pore sizes to enrich pathogens found on fresh-cut cantaloupe...
2016: PloS One
https://www.readbyqxmd.com/read/27852281/viral-bacterial-coinfection-affects-the-presentation-and-alters-the-prognosis-of-severe-community-acquired-pneumonia
#20
Guillaume Voiriot, Benoit Visseaux, Johana Cohen, Liem Binh Luong Nguyen, Mathilde Neuville, Caroline Morbieu, Charles Burdet, Aguila Radjou, François-Xavier Lescure, Roland Smonig, Laurence Armand-Lefèvre, Bruno Mourvillier, Yazdan Yazdanpanah, Jean-Francois Soubirou, Stephane Ruckly, Nadhira Houhou-Fidouh, Jean-François Timsit
BACKGROUND: Multiplex polymerase chain reaction (mPCR) enables recovery of viruses from airways of patients with community-acquired pneumonia (CAP), although their clinical impact remains uncertain. METHODS: Among consecutive adult patients who had undergone a mPCR within 72 hours following their admission to one intensive care unit (ICU), we retrospectively included those with a final diagnosis of CAP. Four etiology groups were clustered: bacterial, viral, mixed (viral-bacterial) and no etiology...
October 25, 2016: Critical Care: the Official Journal of the Critical Care Forum
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