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Multiplex PCR

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https://www.readbyqxmd.com/read/28926935/identification-of-circulating-mirnas-differentially-regulated-by-opioid-treatment
#1
Kaoru Toyama, Naoki Kiyosawa, Kenji Watanabe, Hitoshi Ishizuka
Emerging evidence demonstrates functional contributions of microRNAs (miRNAs) to μ-opioid receptor (MOR) signaling, but the information so far has been mostly limited to their intracellular regulatory mechanisms. The present study aimed to investigate changes in plasma miRNA profiles elicited by opioid treatment in blood samples collected from clinical studies. Healthy male subjects were orally administered with hydromorphone or oxycodone and blood samples were collected at a specified time after the drug treatment...
September 16, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28926641/impact-of-monotherapy-on-hiv-1-reservoir-immune-activation-and-co-infection-with-epstein-barr-virus
#2
Maria Raffaella Petrara, Anna Maria Cattelan, Lolita Sasset, Riccardo Freguja, Francesco Carmona, Silvia Sanavia, Marisa Zanchetta, Paola Del Bianco, Anita De Rossi
OBJECTIVES: Although monotherapy (mART) effectiveness in maintaining viral suppression and CD4 cell count has been extensively examined in HIV-1-infected patients, its impact on HIV-1 reservoir, immune activation, microbial translocation and co-infection with Epstein-Barr Virus (EBV) is unclear. METHODS: This retrospective study involved 32 patients who switched to mART; patients were studied at baseline, 48 and 96 weeks after mART initiation. Thirty-two patients who continued combined antiretroviral therapy (cART) over the same period of time were included in the study...
2017: PloS One
https://www.readbyqxmd.com/read/28925082/prospective-evaluation-of-the-septifast-multiplex-real-time-pcr-assay-for-surveillance-and-diagnosis-of-infections-in-haematological-patients-after-allogeneic-stem-cell-transplantation-compared-to-routine-microbiological-assays-and-an-in-house-real-time-pcr
#3
Sandra Elges, Renate Arnold, Oliver Liesenfeld, Grzegorz Kofla, Agata Mikolajewska, Stefan Schwartz, Lutz Uharek, Markus Ruhnke
We prospectively evaluated a multiplex real-time PCR assay (SeptiFast, SF) in a cohort of patients undergoing allo-BMT in comparison to an in-house PCR method (IH-PCR). Overall 847 blood samples (mean 8 samples/patient) from 104 patients with haematological malignancies were analysed. The majority of patients had acute leukaemia (62%) with a mean age of 52 years (54% female). Pathogens could be detected in 91 of 847 (11%) samples by SF compared to 38 of 205 (18.5%) samples by BC, and 57 of 847 (6.7%) samples by IH-PCR...
September 19, 2017: Mycoses
https://www.readbyqxmd.com/read/28923715/a-multiplex-bead-based-assay-for-immune-gene-expression-analysis-in-shrimp
#4
Sopacha Arayamethakorn, Nitsara Karoonuthaisiri, Wanilada Rungrassamee
Here, we developed a 9-plex bead-based array as a tool to evaluate molecular effects on transcription levels of immune-related genes in the black tiger shrimp (Penaeus monodon). The bead array technology allows simultaneous detection of multiple target genes in a single sample, reducing time, labor and cost. The oligonucleotide probes were designed to target eight immune-related genes that involve in antimicrobial activity, melanization, pathogen pattern recognition proteins, lysozyme and one housekeeping gene as an internal control...
September 15, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/28923133/genetic-evidence-for-a-novel-variant-of-the-pilus-island-1-backbone-protein-in-group-b-streptococcus
#5
Sarah Teatero, Alefiya Neemuchwala, Kunlin Yang, Janice Gomes, Taryn B T Athey, Irene Martin, Walter Demczuk, Allison McGeer, Nahuel Fittipaldi
PURPOSE: Pili contribute significantly to the pathogenesis of infection of group B Streptococcus (GBS) by facilitating adhesion and invasion of host cells. GBS pilin subunits (the backbone pilin protein, BP, and the ancillary pilin proteins, AP) as well as the specific enzymes required for pilus assembly are encoded by genes located in two separate genomic regions, known as pilus island 1 (PI-1) and PI-2. Our aim was to characterize the pilus profile of a collection of GBS isolates from metropolitan Toronto, Canada...
September 19, 2017: Journal of Medical Microbiology
https://www.readbyqxmd.com/read/28921905/antibiotic-resistant-escherichia-coli-in-southeastern-australian-pig-herds-and-implications-for-surveillance
#6
L K van Breda, O P Dhungyel, M P Ward
To investigate public health implications of antibiotics to control post-weaning scours, we surveyed 22 commercial pig herds in southeastern Australia. Fifty faecal samples per herd were collected from pre- and post-weaned piglets. Presumptive Escherichia coli isolates were confirmed by MALDI-TOF MS. Isolates (n = 325) were screened for susceptibility to 19 veterinary antibiotics using MIC broth microdilution. All 325 E. coli isolates underwent further testing against 27 antibiotics used in human medicine and were screened for ETEC adhesin and enterotoxin genes (F4 (K88), F5 (K99), F6 (987P), F18, F41, STa, STb, Stx2e and LT) by multiplex PCR...
September 17, 2017: Zoonoses and Public Health
https://www.readbyqxmd.com/read/28921387/analysis-of-24-genes-reveals-a-monogenic-cause-in-11-1-of-cases-with-steroid-resistant-nephrotic-syndrome-at-a-single-center
#7
Weizhen Tan, Svjetlana Lovric, Shazia Ashraf, Jia Rao, David Schapiro, Merlin Airik, Shirlee Shril, Heon Yung Gee, Michelle Baum, Ghaleb Daouk, Michael A Ferguson, Nancy Rodig, Michael J G Somers, Deborah R Stein, Asaf Vivante, Jillian K Warejko, Eugen Widmeier, Friedhelm Hildebrandt
BACKGROUND: Steroid-resistant nephrotic syndrome (SRNS) is the second most frequent cause of end-stage renal disease (ESRD) among patients manifesting at under 25 years of age. We performed mutation analysis using a high-throughput PCR-based microfluidic technology in 24 single-gene causes of SRNS in a cohort of 72 families, who presented with SRNS before the age of 25 years. METHODS: Within an 18-month interval, we obtained DNA samples, pedigree information, and clinical information from 77 consecutive children with SRNS from 72 different families seen at Boston Children's Hospital (BCH)...
September 18, 2017: Pediatric Nephrology: Journal of the International Pediatric Nephrology Association
https://www.readbyqxmd.com/read/28919697/16s-ribosomal-rna-sequencing-and-molecular-serotyping-of-avibacterium-paragallinarum-isolated-from-indian-field-conditions
#8
Vihang Vithalrao Patil, Debendranath Mishra, Dilip Vithalrao Mane
AIM: This study was aimed at identifying Indian field isolates of Avibacterium paragallinarum on both molecular as well as serological levels that cause infectious coryza in chickens. MATERIALS AND METHODS: Species-specific polymerase chain reaction (HPG-2 PCR), and 16S ribosomal RNA (rRNA) sequencing were employed for molecular identification. Whereas, multiplex PCR technique was used for serological identification of Indian field isolates of A. paragallinarum...
August 2017: Veterinary World
https://www.readbyqxmd.com/read/28919685/prevalence-of-listeria-monocytogenes-yersinia-enterocolitica-staphylococcusaureus-and-salmonella-enterica-typhimurium-in-meat-and-meat-products-using-multiplex-polymerase-chain-reaction
#9
C Latha, C J Anu, V J Ajaykumar, B Sunil
AIM: The objective of the study was to investigate the occurrence of Listeria monocytogenes, Yersinia enterocolitica, Staphylococcusaureus, and Salmonella enterica Typhimurium in meat and meat products using the multiplex polymerase chain reaction (PCR) method. MATERIALS AND METHODS: The assay combined an enrichment step in tryptic soy broth with yeast extract formulated for the simultaneous growth of target pathogens, DNA isolation and multiplex PCR. A total of 1134 samples including beef (n=349), chicken (n=325), pork (n=310), chevon (n=50), and meat products (n=100) were collected from different parts of Kerala, India...
August 2017: Veterinary World
https://www.readbyqxmd.com/read/28919671/detection-of-staphylococcus-aureus-enterotoxigenic-strains-in-bovine-raw-milk-by-reversed-passive-latex-agglutination-and-multiplex-polymerase-chain-reaction
#10
Asmaa Samy Mansour, Gad El-Said Wagih, Sabry D Morgan, Mahmoud Elhariri, Mona A El-Shabrawy, Azza S M Abuelnaga, E A Elgabry
AIM: This review gives an outline of the assessment of enterotoxigenic Staphylococcus aureus tainting levels in raw milk from different sources in Egypt and characterization of enterotoxigenic strains utilizing a technique in light of PCR to identify genes coding for the production of staphylococcal enterotoxin (SE). The obtained data were compared with results from the application of the reversed passive latex. MATERIALS AND METHODS: Multiplex PCR and reversed passive latex agglutination (RPLA) were used...
August 2017: Veterinary World
https://www.readbyqxmd.com/read/28919618/molecular-detection-and-differentiation-of-mycobacterium-tuberculosis-complex-and-non-tuberculous-mycobacterium-in-the-clinical-specimens-by-real-time-pcr
#11
M M Rahman, M R Rahim, A Khaled, T A Nasir, F Nasrin, M A Hasan
Mycobacteria are subdivided into three groups: the Mycobacterium tuberculosis complex, the non-tuberculous mycobacteria called NTM or MOTT (Mycobacteria Other Than Tuberculosis) and Mycobacterium leprae. Over the past few decades, the incidence of infections caused by NTM has increased world wide. The differentiation of Mycobacterium tuberculosis complex from NTM is of primary importance for infection control and choice of antimicrobial therapy. However, there is so far no report in Bangladesh about the detection of NTM and hence differentiation of MTB and NTM...
July 2017: Mymensingh Medical Journal: MMJ
https://www.readbyqxmd.com/read/28919163/multiplex-pcr-and-ngs-based-identification-of-mrna-splicing-variants-analysis-of-brca1-splicing-pattern-as-a-model
#12
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...
September 14, 2017: Gene
https://www.readbyqxmd.com/read/28918548/tumor-and-serum-dna-methylation-in-women-receiving-preoperative-chemotherapy-with-or-without-vorinostat-in-tbcrc008
#13
Roisin M Connolly, Mary Jo Fackler, Zhe Zhang, Xian C Zhou, Matthew P Goetz, Judy C Boughey, Bridget Walsh, John T Carpenter, Anna Maria Storniolo, Stanley P Watkins, Edward W Gabrielson, Vered Stearns, Saraswati Sukumar
BACKGROUND: Methylated gene markers have shown promise in predicting breast cancer outcomes and treatment response. We evaluated whether baseline and changes in tissue and serum methylation levels would predict pathological complete response (pCR) in patients with HER2-negative early breast cancer undergoing preoperative chemotherapy. METHODS: The TBCRC008 trial investigated pCR following 12 weeks of preoperative carboplatin and albumin-bound paclitaxel + vorinostat/placebo (n = 62)...
September 16, 2017: Breast Cancer Research and Treatment
https://www.readbyqxmd.com/read/28918076/simultaneous-detection-of-wheat-dwarf-virus-northern-cereal-mosaic-virus-barley-yellow-striate-mosaic-virus-and-rice-black-streaked-dwarf-virus-in-wheat-by-multiplex-rt-pcr
#14
Peipei Zhang, Yan Liu, Wenwen Liu, Sebastien Massart, Xifeng Wang
Wheat dwarf virus (WDV), barley yellow striate mosaic virus (BYSMV), rice black-streaked dwarf virus (RBSDV) and northern cereal mosaic virus (NCMV) are four viruses infecting wheat and causing similar symptoms. In this paper, a multiplex reverse transcription polymerase chain reaction (m-RT-PCR) method has been developed for the simultaneous detection and discrimination of these viruses. The protocol uses specific primer set for each virus and produces four distinct fragments (273, 565, 783 and 1296bp), detecting the presence of RBSDV, BYSMV, WDV and NCMV, respectively...
September 13, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28916427/simultaneous-detection-and-differentiation-of-three-potyviridae-viruses-in-sweet-potato-by-a-multiplex-taqman-real-time-rt-pcr-assay
#15
Pingxiu Lan, Fan Li, Jorge Abad, Lingling Pu, Ruhui Li
A multiplex TaqMan real time RT-PCR was developed for detection and differentiation of Sweet potato virus G, Sweet potato latent virus and Sweet potato mild mottle virus in one tube. Amplification and detection of a fluorogenic cytochrome oxidase gene was included as an internal control. The assay was compared with a multiplex RT-PCR developed in the initial study for the detection and differentiation of the three viruses and host 18S rRNA. Primers and/or probes of the two assays were designed from conserved regions of each virus...
September 12, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28916184/role-of-the-il-12-il-35-balance-in-sj%C3%A3-gren-s-syndrome
#16
Olivier Fogel, Elodie Rivière, Raphaèle Seror, Gaetane Nocturne, Saida Boudaoud, Bineta Ly, Jacques-Eric Gottenberg, Véronique Le Guern, Jean-Jacques Dubost, Joanne Nititham, Kimberly E Taylor, Philippe Chanson, Philippe Dieudé, Lindsey A Criswell, Bernd Jagla, Alice Thai, Michael Mingueneau, Xavier Mariette, Corinne Miceli-Richard
BACKGROUND: An interferon (IFN) signature is involved in the pathogenesis of primary Sjögren's syndrome (pSS), but whether the signature is type 1 or 2 remains controversial. Mouse models and genetic studies suggested the involvement of T helper 1 and type 2 IFN pathways. Likewise, polymorphisms of interleukin 12A gene (IL-12A), which encodes for IL-12p35, have been associated with pSS. IL-12p35 subunit is shared by 2 heterodimers, IL-12 and IL-35. OBJECTIVE: To confirm the genetic association of IL-12A polymorphism and pSS and elucidate the involvement of the IL-12/IL-35 balance in pSS by functional studies...
September 12, 2017: Journal of Allergy and Clinical Immunology
https://www.readbyqxmd.com/read/28916034/colorimetric-biosensor-based-on-a-dnazyme-primer-and-its-application-in-logic-gate-operations-for-dna-screening
#17
Chenguang Wang, Nan Cheng, Longjiao Zhu, Yuancong Xu, Kunlun Huang, Pengyu Zhu, Shuifang Zhu, Wei Fu, Wentao Xu
A colorimetric biosensor for DNA screening was designed based on the conformational changes of the horseradish peroxidase (HRP)-mimicking DNAzyme. The scheme of DNA biosensing was designed based on the base pairing of DNAzyme sequence to inhibit the formation of HRP-mimicking hemin/G-quadruplex structures in the process of amplification. DNA could be amplified via the universal primer multiplex polymerase chain reaction (UP-M-PCR) and innovatively detected as color disappear in the reaction visible to the naked eye...
September 22, 2017: Analytica Chimica Acta
https://www.readbyqxmd.com/read/28915725/distribution-of-rotavirus-genotypes-ccirculating-in-ahvaz-iran-in-2016
#18
Azarakhsh Azaran, Manoochehr Makvandi, Ali Teimoori, Saeedeh Ebrahimi, Farzad Heydari, Roya Nikfar
Background: Group A rotavirus (RVA) mainly causes acute gastroenteritis, exclusively in young children in developing countries. The prevalence and determination of the molecular epidemiology of rotavirus genotypes will determine the dominant rotavirus genotypes in the region and provide a strategy for the development of appropriate vaccines. Methods: A total of 100 fecal samples were collected from children below five years with acute gastroenteritis who referred to Aboozar Children’s Hospital of Ahvaz city during October 2015 to March 2016...
September 16, 2017: Iranian Biomedical Journal
https://www.readbyqxmd.com/read/28915607/coal-tar-pitch-extract-could-induce-chromosomal-instability-of-human-bronchial-epithelial-cells-mediated-by-spindle-checkpoint-related-proteins
#19
Peng Zhang, Zhitao Li, Na Wang, Guangcai Duan, Wei Wang, Yanming Feng, Yong Zhao, Lixia Wang, Hansong Zhu, Qiao Zhang, Xiaozhuan Liu, Weidong Wu, Yongjun Wu, Wu Yao, Jing Wang, Yiming Wu, Feifei Feng
Coal tar pitch (CTP) is a byproduct of coal tar distillation. The workers working with coal tar or in aluminum smelters, potrooms and carbon plants have the opportunities of exposing to coal tar pitch volatiles. Coal tar pitches from which polycyclic aromatic hydrocarbons (PAHs) originate have been shown to exhibit lung carcinogenicity in humans. Chromosomal instability (CIN) is a mechanism in carcinogenesis, however, whether CIN is involved in coal tar pitch-induced lung cancer remains elusive. In this present study, human bronchial epithelial cells (BEAS-2B) were first exposed to coal tar pitch extracts (CTPE) to induce a malignant transformation model...
August 22, 2017: Oncotarget
https://www.readbyqxmd.com/read/28911843/the-effect-of-a-cannula-milk-sampling-technique-on-the-microbiological-diagnosis-of-bovine-mastitis
#20
M Friman, H Hiitiö, M Niemi, J Holopainen, S Pyörälä, H Simojoki
Two methods of collecting milk samples from mastitic bovine mammary quarters were compared. Samples were taken in a consistent order in which standard aseptic technique sampling was done first, followed by insertion of a sterile cannula through the teat canal and collection of a second sample. Microbiological results of those two sampling techniques were compared. Milk samples were analysed using multiplex real-time polymerase chain reaction (PCR). The cannula technique produced a reduced number of microbial species or groups of species per sample compared with conventional sampling...
August 2017: Veterinary Journal
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