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Rapid PCR

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https://www.readbyqxmd.com/read/28526036/molecular-characterization-of-dirofilaria-spp-circulating-in-portugal
#1
Cátia Ferreira, Ana Afonso, Manuela Calado, Isabel Maurício, Ana Margarida Alho, José Meireles, Luís Madeira de Carvalho, Silvana Belo
BACKGROUND: Dirofilariosis is a potentially zoonotic parasitic disease, mainly transmitted by mosquito vectors in many parts of the world. Data concerning the canine Dirofilaria species currently circulating in Portugal is scarce. Thereby, a large-scale study was conducted to determine the Dirofilaria spp. present in Portugal, based on a molecular approach, and also to optimize a reliable and highly sensitive species-specific polymerase chain reaction (PCR) assay that could be used for the simultaneous detection and differentiation of Dirofilaria immitis, Dirofilaria repens, and other concurrent filarial species in animal reservoirs...
May 19, 2017: Parasites & Vectors
https://www.readbyqxmd.com/read/28525954/construction-of-various-%C3%AE-34-5-deleted-fluorescent-expressing-oncolytic-herpes-simplex-type-1-ohsv-for-generation-and-isolation-of-hsv-based-vectors
#2
REVIEW
Shahriyar Abdoli, Farzin Roohvand, Ladan Teimoori-Toolabi, Mohammad Ali Shokrgozar, Mina Bahrololoumi, Kayhan Azadmanesh
Background: Oncolytic herpes simplex virus (oHSV)-based vectors lacking γ34.5 gene, are considered as ideal templates to construct efficient vectors for (targeted) cancer gene therapy. Herein, we reported the construction of three single/dually-flourescence labeled and γ34.5-deleted, recombinant HSV-1 vectors for rapid generation and easy selection/isolation of different HSV-Based vectors. Methods: Generation of recombinant viruses was performed with conventional homologous recombination methods using green fluorescent protein (GFP) and BleCherry harboring shuttle vectors...
July 2017: Iranian Biomedical Journal
https://www.readbyqxmd.com/read/28525658/genotyping-of-snps-associated-with-meat-tenderness-comparison-of-two-pcr-based-methods
#3
L E López-Rojas, L Patiño-Cadavid, A López-Herrera, J J Echeverri-Zuluaga
Single nucleotide polymorphisms (SNPs) carried in calpain (CAPN1), calpastatin (CAST), and leptin (LEP) genes are associated with meat tenderness. Due to the economic importance of this meat quality attribute, the development of fast, reliable, and affordable methods to identify bovine carriers of favorable alleles is of great importance for genetic improvement. Currently, PCR-RFLP is accepted as the standard gold method for genotyping SNPs associated with meat tenderness. But these SNPs can be detected by other techniques as high-resolution melting (HRM) analysis - a post-PCR method - that offers several advantages and has great application potential in the meat industry...
May 18, 2017: Genetics and Molecular Research: GMR
https://www.readbyqxmd.com/read/28525340/detection-of-bordetella-avium-by-taqman-real-time-pcr-in-tracheal-swabs-from-wildlife-birds
#4
T Stenzel, D Pestka, B Tykałowski, M Śmiałek, A Koncicki, A Bancerz-Kisiel
Bordetella avium, the causing agent of bordetellosis, a highly contagious infection of the respiratory tract in young poultry, causes significant losses in poultry farming throughout the world. Wildlife birds can be a reservoir of various pathogens that infect farm animals. For this reason the studies were conducted to estimate the prevalence of Bordetella avium in wildlife birds in Poland. Tracheal swab samples were collected from 650 birds representing 27 species. The bacterial DNA was isolated directly from the swabs and screened for Bordetella avium by TaqMan real-time PCR...
March 28, 2017: Polish Journal of Veterinary Sciences
https://www.readbyqxmd.com/read/28522856/diagnosing-congenital-malaria-in-a-high-transmission-setting-clinical-relevance-and-usefulness-of-p-falciparum-hrp2-based-testing
#5
Hamtandi Magloire Natama, Delwendé Florence Ouedraogo, Hermann Sorgho, Eduard Rovira-Vallbona, Elisa Serra-Casas, M Athanase Somé, Maminata Coulibaly-Traoré, Petra F Mens, Luc Kestens, Halidou Tinto, Anna Rosanas-Urgell
Congenital malaria diagnosis is challenging due to frequently observed low parasite density infections, while their clinical relevance during early infancy is not well characterized. In Nanoro health district (Burkina Faso), we determined the prevalence of congenital malaria by real-time quantitative PCR and we assessed the performance of rapid diagnosis test (RDT) and light microscopy (LM) to detect Plasmodium falciparum infections in cord-blood samples. In addition, we examined the usefulness of P. falciparum Histidine Rich Protein2 (PfHRP2) as surrogate biomarker of infection and explored association between congenital malaria and clinical outcomes...
May 18, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28522818/efficient-modification-of-%C3%AE-dna-substrates-for-single-molecule-studies
#6
Yoori Kim, Armando de la Torre, Andrew A Leal, Ilya J Finkelstein
Single-molecule studies of protein-nucleic acid interactions frequently require site-specific modification of long DNA substrates. The bacteriophage λ is a convenient source of high quality long (48.5 kb) DNA. However, introducing specific sequences, tertiary structures, and chemical modifications into λ-DNA remains technically challenging. Most current approaches rely on multi-step ligations with low yields and incomplete products. Here, we describe a molecular toolkit for rapid preparation of modified λ-DNA...
May 18, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28522225/genetic-screening-of-spinal-muscular-atrophy-using-a-real-time-modified-cop-pcr-technique-with-dried-blood-spot-dna
#7
Mawaddah Ar Rochmah, Nur Imma Fatimah Harahap, Emma Tabe Eko Niba, Kenta Nakanishi, Hiroyuki Awano, Ichiro Morioka, Kazumoto Iijima, Toshio Saito, Kayoko Saito, Poh San Lai, Yasuhiro Takeshima, Atsuko Takeuchi, Yoshihiro Bouike, Maya Okamoto, Hisahide Nishio, Masakazu Shinohara
BACKGROUND: Spinal muscular atrophy (SMA) is a common neuromuscular disorder caused by mutations in SMN1. More than 95% of SMA patients carry homozygous SMN1 deletion. SMA is the leading genetic cause of infant death, and has been considered an incurable disease. However, a recent clinical trial with an antisense oligonucleotide drug has shown encouraging clinical efficacy. Thus, early and accurate detection of SMN1 deletion may improve prognosis of many infantile SMA patients. METHODS: A total of 88 DNA samples (37 SMA patients, 12 carriers and 39 controls) from dried blood spots (DBS) on filter paper were analyzed...
May 15, 2017: Brain & Development
https://www.readbyqxmd.com/read/28521058/enhanced-expression-of-proteins-involved-in-energy-production-and-transfer-in-breast-muscle-of-pedigree-male-broilers-exhibiting-high-feed-efficiency
#8
W G Bottje, K Lassiter, S Dridi, N Hudson, B-W Kong
In cells with fluctuating energy demand (e.g., skeletal muscle), a transfer system of proteins across the inner and outer mitochondrial membranes links mitochondrial oxidative phosphorylation to cytosolic phosphorylated creatine (PCr) that serves as a phosphate reservoir for rapid repletion of cytosolic adenosine triphosphate (ATP). Crucial proteins of this energy transfer system include several creatine kinase (CK) isoforms found in the cytosol and mitochondria. In a recent proteomic study (Kong et al., 2016), several components of this system were up-regulated in high feed efficiency (FE) compared to low FE breast muscle; notably adenine nucleotide translocase (ANT), voltage dependent activated channel (VDAC), the brain isoform of creatine kinase (CK-B), and several proteins of the electron transport chain...
May 17, 2017: Poultry Science
https://www.readbyqxmd.com/read/28520858/rapid-tests-for-influenza-respiratory-syncytial-virus-and-other-respiratory-viruses-a-systematic-review-and-meta-analysis
#9
Ahl Bruning, Mmg Leeflang, Jmbw Vos, R Spijker, M D de Jong, K C Wolthers, D Pajkrt
Rapid diagnosis of respiratory virus infections contributes to patient care. This systematic review evaluates the diagnostic accuracy of rapid tests for the detection of respiratory viruses. We searched Medline and EMBASE for studies evaluating these tests against PCR as reference standard. 179 studies were included of which 134 evaluated rapid tests for influenza viruses, 32 for RSV, and 13 for other respiratory viruses. We used the bivariate random effects model for quantitative meta-analysis of the results...
May 17, 2017: Clinical Infectious Diseases: An Official Publication of the Infectious Diseases Society of America
https://www.readbyqxmd.com/read/28520818/evaluation-of-a-real-time-method-of-simultaneous-amplification-and-testing-in-diagnosis-of-mycoplasma-pneumoniae-infection-in-children-with-pneumonia
#10
Wei Li, You-Hong Fang, Hong-Qiang Shen, De-Hua Yang, Qiang Shu, Shi-Qiang Shang
Mycoplasma pneumoniae (M. pneumoniae) infection can cause community acquired pneumonia in children. A real-time method of simultaneous amplification and testing of M. pneumoniae (SAT-MP) was developed to diagnose M. pneumoniae targeting a region of the ribosomal RNA. The SAT-MP assay can accurately identify M. pneumoniae with a detection range from 101 to 107 CFU/ml. In this study, the specimens from 315 children with pneumonia were collected and analyzed by SAT-MP in parallel with real-time PCR method and IgM ELISA assay...
2017: PloS One
https://www.readbyqxmd.com/read/28516136/primers-and-probe-design-and-precision-assessment-of-the-real-time-rt-pcr-assay-in-coxsackievirus-a10-and-enterovirus-detection
#11
Jingfang Chen, Rusheng Zhang, Xinhua Ou, Dong Yao, Zheng Huang, Linzhi Li, Biancheng Sun
This data article contains data related to the research article entitled "Rapid detection of enterovirus and Coxsackievirus A10 by a TaqMan based duplex one-step real time RT-PCR assay" (Chen at al., 2017) [1]. Primers and probe sequence design are among the most critical factors in real-time polymerase chain reaction (PCR) assay optimization. Linearity, sensitivity, specificity and precision are the crucial criteria which are used to evaluate the performance of a new method. This data article report the primers and probe design and precision assessment of the new assay...
June 2017: Data in Brief
https://www.readbyqxmd.com/read/28515220/culture-based-methods-and-molecular-tools-for-azole-resistant-aspergillus-fumigatus-detection-in-a-belgian-university-hospital
#12
I Montesinos, M A Argudín, M Hites, F Ahajjam, M Dodémont, C Dagyaran, M Bakkali, I Etienne, F Jacobs, C Knoop, S Patteet, K Lagrou
Azole-resistant Aspergillus fumigatus is an increasing worldwide problem with major clinical implications. Surveillance is warranted to guide clinicians to provide optimal treatment to patients. To investigate azole resistance in clinical Aspergillus isolates in our institution, a Belgian university hospital, we conducted a laboratory-based surveillance between June 2015 and October 2016. Two different approaches were used: a prospective culture-based surveillance using VIPcheck™ on unselected A. fumigatus (n=109 patients, including 19 patients with proven or probable IA), followed by molecular detection of mutations conferring azole resistance and a retrospective detection of azole-resistant A...
May 17, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28515219/a-novel-stool-pcr-test-for-helicobacter-pylori-may-predict-clarithromycin-resistance-and-patient-eradication-at-a-high-rate
#13
Erin Beckman, Ilaria Saracino, Giulia Fiorini, Courtney Clark, Vladimir Slepnev, Denise Patel, Clarissa Gomez, Reddy Ponaka, Vecheslav Elagin, Dino Vaira
Objectives: Clarithromycin-based regimens are commonly used as a first line therapy in H. pylori positive patients, however treatment failures are associated with resistance to clarithromycin. The aim of this study was to evaluate the feasibility of using stool samples to detect the presence of H. pylori DNA, while concurrently detecting mutations associated with the resistance to clarithromycin.Methods: Total DNA was extracted from 294 raw stools of H pylori positive and negative patients. TaqMan real-time PCR amplification was used to detect the presence of H...
May 17, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28515213/rapid-identification-of-five-classes-of-carbapenem-resistance-genes-directly-from-rectal-swabs-using-the-xpert%C3%A2-carba-r-assay
#14
Nicholas M Moore, Rafael Cantón, Edoardo Carretto, Lance R Peterson, Robert L Sautter, Maria M Traczewski
Carbapenemase-producing organisms (CPO) have been identified by global health leaders as an urgent threat. Detection of patients with gastrointestinal carriage of CPO is necessary to interrupt their spread within healthcare facilities. In this multi-site study, we assessed the performance of the Xpert® Carba-R test, a rapid real-time PCR (qPCR) assay that detects five families of carbapenemase genes (blaIMP, blaKPC, blaNDM, blaOXA-48, and blaVIM) directly from rectal swab specimens. Using dual swabs, specimens from 755 patients were collected and tested prospectively...
May 17, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28515100/detection-of-chromosomal-translocation-in-hematologic-malignancies-by-a-novel-dna-based-looped-ligation-assay-lola
#15
Shuko Harada, Emily Sizzle, Ming-Tseh Lin, Christopher D Gocke
BACKGROUND: Disease-defining chromosomal translocations are seen in various neoplasms, especially in lymphomas and leukemias. Translocation detection at the DNA level is often complicated by chromosomal breakpoints that are distributed over very large regions. We have developed a ligation-based assay [the looped ligation assay (LOLA)] to detect translocations from diseases with multiple widely spaced breakpoint hot spots. METHODS: Oligonucleotide sets that probe breakpoints of IGH-BCL2 (immunoglobulin heavy-apoptosis regulator) in follicular lymphoma (FL), MYC-IGH (MYC proto-oncogene, bHLH transcription factor- immunoglobulin heavy) in Burkitt lymphoma (BL) and BCR-ABL1 (RhoGEF and GTPase activating protein-ABL proto-oncogene 1, non-receptor tyrosine kinase) in chronic myelogenous leukemia (CML) were designed...
May 17, 2017: Clinical Chemistry
https://www.readbyqxmd.com/read/28513759/-diagnosis-of-leptospirosis-in-dogs
#16
Michèle Bergmann, Julia-Rebecca Llewellyn, Katrin Hartmann
Rapid confirmation of the diagnosis leptospirosis is important in order to apply appropriate treatment; in addition, infected dogs are a zoonotic risk. Culture and isolation of living leptospires from blood, urine, or tissues is considered the reference standard of diagnosis. However, sensitivity are low, and leptospires require weeks to months to grow. Therefore, diagnosis of leptospirosis is most commonly based on antibody testing or the detection of Leptospira spp. DNA by PCR. Microscopic agglutination test (MAT) is currently still the recommended confirmatory test for canine leptospirosis, despite its numerous limitations (e...
May 17, 2017: Tierärztliche Praxis. Ausgabe K, Kleintiere/Heimtiere
https://www.readbyqxmd.com/read/28513665/a-fully-integrated-microchip-system-for-automated-forensic-short-tandem-repeat-analysis
#17
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/28513282/zanthoxylum-alkylamides-activate-phosphorylated-ampk-and-ameliorate-glycolipid-metabolism-in-the-streptozotocin-induced-diabetic-rats
#18
Tingyuan Ren, Yuping Zhu, Jianquan Kan
This study aimed to evaluate the effects of Zanthoxylum alkylamides on the glycolipid metabolism of rats with streptozotocin (STZ)-induced diabetes. Diabetic rats were given daily oral treatments of 2, 4, or 8 mg/kg bw alkylamides for 28 days. Alkylamides significantly decreased fasting blood glucose and fructosamine content, as well as relieved organ enlargement caused by diabetes. The serum and liver triglyceride, malondialdehyde, and free fatty-acid contents of rats with STZ-induced diabetes were significantly reduced...
May 17, 2017: Clinical and Experimental Hypertension: CHE
https://www.readbyqxmd.com/read/28512861/rapid-diagnosis-of-sepsis-with-taqman-based-multiplex-real-time-pcr
#19
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/28512020/molecular-epidemiology-of-porcine-reproductive-and-respiratory-syndrome-virus-in-central-china-since-2014-the-prevalence-of-nadc30-like-prrsvs
#20
Lin-Jian Wang, Weitao Xie, Xin-Xin Chen, Songlin Qiao, Mengmeng Zhao, Yu Gu, Bao-Lei Zhao, Gaiping Zhang
Porcine reproductive and respiratory syndrome (PRRS), characterized by respiratory disorders in piglets and reproductive failure in sows, is still the great threat of swine industry. Recently, Emergence of the novel NADC30-like PRRS viruses (PRRSVs) has caused widespread outbreaks of PRRS. To investigate the epidemic characteristics of PRRSVs in Central China since 2014, 6372 clinical serum samples were tested by ELISA, 250 tissue samples were tested by RT-PCR, and among these, 30 ORF5 and 17 Nsp2 genes sequences were analyzed...
May 13, 2017: Microbial Pathogenesis
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