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https://www.readbyqxmd.com/read/28433811/rapid-and-sensitive-detection-of-salmonid-alphavirus-using-taqman-real-time-pcr
#1
Wen Shi, Aochen Song, Shuai Gao, Yuting Wang, Lijie Tang, Yigang Xu, Tong Ren, Yijing Li, Min Liu
Salmonid alphavirus (SAV) infection has led to the spread of salmon pancreas disease (PD) and sleeping disease (SD) to salmonids in several countries in Europe, resulting in tremendous economic losses to the fish farming industry. Recently, with increases in the fish import trade, many countries in which SAV has been unreported, such as China, may be seriously threatened by these diseases. It is therefore necessary to develop efficient detection methods for the prevention and diagnosis of SAV infection. In this study, a rapid and sensitive TaqMan real-time PCR method was established and assessed for this purpose...
April 19, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28433579/distinguishing-septic-from-normal-donors-by-detection-of-spla2-iia-from-human-plasma-using-a-microsieve-based-immunoassay
#2
Daniel R Zweitzig, Arjan G Tibbe, Mark J Kopnitsky, Kathleen Cichonski, Leon W M M Terstappen
Bloodstream infections that progress to septic shock are responsible for hundreds of thousands of deaths each year, and are associated with significant healthcare costs. Recent studies have shown that a member of the secreted phospholipase protein family, termed sPLA2-IIA, may play a role during the innate immune response to bacterial infections, and is elevated in the plasma of septic patients. In this report, the feasibility of a simple microsieve-based sPLA2-IIA detection immunoassay was explored. Microsieves containing 5μm pores were covalently coupled with a sPLA2-IIA-specific monoclonal antibody at 0...
April 19, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28428345/proliferation-of-primary-human-hepatocytes-and-prevention-of-hepatitis-b-virus-reinfection-efficiently-deplete-nuclear-cccdna-in-vivo
#3
Lena Allweiss, Tassilo Volz, Katja Giersch, Janine Kah, Giuseppina Raffa, Joerg Petersen, Ansgar W Lohse, Concetta Beninati, Teresa Pollicino, Stephan Urban, Marc Lütgehetmann, Maura Dandri
OBJECTIVE: The stability of the covalently closed circular DNA (cccDNA) in nuclei of non-dividing hepatocytes represents a key determinant of HBV persistence. Contrarily, studies with animal hepadnaviruses indicated that hepatocyte turnover can reduce cccDNA loads but knowledge on the proliferative capacity of HBV-infected primary human hepatocytes (PHHs) in vivo and the fate of cccDNA in dividing PHHs is still lacking. This study aimed to determine the impact of human hepatocyte division on cccDNA stability in vivo...
April 20, 2017: Gut
https://www.readbyqxmd.com/read/28424786/immunocytochemistry-improving-the-diagnosis-of-trichomonas-vaginalis-infections
#4
Thaisa Helena S Fonseca, Fabrício M Silva Oliveira, Marina Alacoque, Marcella Israel Rocha, Henrique Vitor Leite, Joseph F Guimarães Santos, Haendel G N O Busatti, Marcelo Vidigal Caliari, Maria Aparecida Gomes
The aim of this study was to evaluate the immunocytochemistry (ICC) to diagnose trichomoniasis, particularly asymptomatic infections. By culture serial dilutions, ICC was able to detect 1 trophozoite/mL, while the culture was positive up to 100 trophozoites/mL. The ICC in vivo detection capability was assessed in vaginal secretions of mice experimentally infected and in vaginal swabs from asymptomatic HIV-positive pregnant women compared with culture. All vaginal secretion samples from mice were positive according to both methods...
2017: BioMed Research International
https://www.readbyqxmd.com/read/28424022/analytical-validation-of-an-automated-assay-for-ferric-reducing-ability-of-plasma-in-dog-serum
#5
Camila Peres Rubio, Silvia Martinez-Subiela, Josefa Hernández-Ruiz, Asta Tvarijonaviciute, José Joaquín Ceron
We performed analytical validation of an automated ferric-reducing ability of plasma (FRAP) assay in the serum of dogs. Intra- and interassay precision, accuracy, detection limit, and effects of hemolysis and lipemia were evaluated. Intra- and interassay coefficients of variation were <1% and <13%, respectively. The assay showed a high correlation with a FRAP assay described previously, and results were linear when serial sample dilutions were tested. The detection limit was lower than the values observed in sera from healthy dogs; decreased serum FRAP was found in dogs with leishmaniosis...
April 1, 2017: Journal of Veterinary Diagnostic Investigation
https://www.readbyqxmd.com/read/28417624/cow-s-milk-desensitization-in-anaphylactic-patients-a-new-personalized-dose-method
#6
Delara Babaie, Mohammad Nabavi, Saba Arshi, Mehrnaz Mesdaghi, Zahra Chavoshzadeh, Mohammad Hasan Bemanian, Mitra Tafakori, Mehrdad Amirmoini, Hosein Esmailzadeh, Rasoul Molatefi, Mahsa Rekabi, Nadieh Akbarpour, Farimah Masoumi, Morteza Fallahpour
Cow's milk allergy (CMA) is the most frequent food allergy in children and oral immunotherapy (OIT) is a promising approach for treatment of patients. The most challenging cases are anaphylactic with coexisting asthma and proposing safe protocols is crucial especially in high risk groups. Considering that CMA varies among patients, an individualized OIT protocol would be beneficial to achieve a safer and more efficient method of desensitization. 18 children more than 3 years of age with IgE-mediated CMA were enrolled...
February 2017: Iranian Journal of Allergy, Asthma, and Immunology
https://www.readbyqxmd.com/read/28414163/droplet-digital-pcr-ddpcr-vs-quantitative-real-time-pcr-qpcr-approach-for-detection-and-quantification-of-merkel-cell-polyomavirus-mcpyv-dna-in-formalin-fixed-paraffin-embedded-ffpe-cutaneous-biopsies
#7
Rosaria Arvia, Mauro Sollai, Federica Pierucci, Carmelo Urso, Daniela Massi, Krystyna Zakrzewska
BACKGROUND: Merkel cell polyomavirus (MCPyV) is associated with Merkel cell carcinoma and high viral load in the skin was proposed as a risk factor for the occurrence of this tumour. MCPyV DNA was detected, with lower frequency, in different skin cancers but since the viral load was usually low, the real prevalence of viral DNA could be underestimated. OBJECTIVE: To evaluate the performance of two assays (qPCR and ddPCR) for MCPyV detection and quantification in formalin fixed paraffin embedded (FFPE) tissue samples...
April 14, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28414024/novel-high-throughput-cell-based-hybridoma-screening-methodology-using-the-celigo-image-cytometer
#8
Haohai Zhang, Leo Li-Ying Chan, William Rice, Nasim Kassam, Maria Serena Longhi, Haitao Zhao, Simon C Robson, Wenda Gao, Yan Wu
Hybridoma screening is a critical step for antibody discovery, which necessitates prompt identification of potential clones from hundreds to thousands of hybridoma cultures against the desired immunogen. Technical issues associated with ELISA- and flow cytometry-based screening limit accuracy and diminish high-throughput capability, increasing time and cost. Conventional ELISA screening with coated antigen is also impractical for difficult-to-express hydrophobic membrane antigens or multi-chain protein complexes...
April 14, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28408937/detection-of-gastrointestinal-pathogens-from-stool-samples-on-hemoccult-cards-by-multiplex-pcr
#9
Martin Alberer, Nicklas Schlenker, Malkin Bauer, Kerstin Helfrich, Carolin Mengele, Thomas Löscher, Hans Dieter Nothdurft, Gisela Bretzel, Marcus Beissner
Purpose. Up to 30% of international travelers are affected by travelers' diarrhea (TD). Reliable data on the etiology of TD is lacking. Sufficient laboratory capacity at travel destinations is often unavailable and transporting conventional stool samples to the home country is inconvenient. We evaluated the use of Hemoccult cards for stool sampling combined with a multiplex PCR for the detection of model viral, bacterial, and protozoal TD pathogens. Methods. Following the creation of serial dilutions for each model pathogen, last positive dilution steps (LPDs) and thereof calculated last positive sample concentrations (LPCs) were compared between conventional stool samples and card samples...
2017: Canadian Journal of Infectious Diseases & Medical Microbiology
https://www.readbyqxmd.com/read/28406722/development-of-quantitative-monoclonal-antibody-specific-immobilization-of-platelet-antigens-assay-for-antibodies-against-human-platelet-antigen-1a-3a-and-5b
#10
R S Li, B Ling, P Lu
Human platelet antigens (HPAs) are platelet-specific alloantigens that cause alloimmunization. Alloantibodies induced via pregnancy and transfusion may result in various clinical syndromes. The monoclonal antibody-specific immobilization of platelet antigens (MAIPA) assay is the gold standard for the detection and identification of HPA antibodies. The present study aimed to develop standardized quantitative MAIPA assays for antibodies against HPA-1a, 3a, and 5b. MAIPA protocol was applied on serial dilutions of standard or reference reagents containing antibodies against HPA-1a, 3a, and 5b...
April 13, 2017: Platelets
https://www.readbyqxmd.com/read/28405178/utilizing-bd-max%C3%A2-enteric-bacterial-panel-to-detect-stool-pathogens-from-rectal-swabs
#11
Barbara DeBurger, Sarah Hanna, Eleanor A Powell, Cindi Ventrola, Joel E Mortensen
BACKGROUND: The BD MAX™ Enteric Bacterial Panel (BDM-EBP) is designed and FDA-cleared to detect Salmonella, Shigella, Campylobacter, and Shiga toxin genes stx1/2 from stool samples. However, rectal swabs, which are not FDA-cleared for clinical testing with the BDM-EBP, are common specimens received from pediatric patients for enteric pathogen testing. The purpose of this study was to evaluate the ability of the BDM-EBP to detect stool pathogens from rectal swabs. METHODS: Routine cultures, Shiga toxin testing, and molecular testing with BDM-EBP were performed on 272 sequential rectal swabs collected from August 2015 to December 2015...
2017: BMC Clinical Pathology
https://www.readbyqxmd.com/read/28405141/effect-of-a-poloxamer-407-based-thermosensitive-gel-on-minimization-of-thermal-injury-to-diaphragm-during-microwave-ablation-of-the-liver
#12
Li-Li Zhang, Gui-Min Xia, Yu-Jiang Liu, Rui Dou, John Eisenbrey, Ji-Bin Liu, Xiao-Wei Wang, Lin-Xue Qian
AIM: To assess the insulating effect of a poloxamer 407 (P407)-based gel during microwave ablation of liver adjacent to the diaphragm. METHODS: We prepared serial dilutions of P407, and 22.5% (w/w) concentration was identified as suitable for ablation procedures. Subsequently, microwave ablations were performed on the livers of 24 rabbits (gel, saline, control groups, n = 8 in each). The P407 solution and 0.9% normal saline were injected into the potential space between the diaphragm and liver in experimental groups...
March 28, 2017: World Journal of Gastroenterology: WJG
https://www.readbyqxmd.com/read/28402500/evaluation-of-the-aid-carbapenemase-line-probe-assay-for-rapid-detection-and-identification-of-carbapenemase-genes-in-gram-negative-bacilli
#13
Guido V Bloemberg, Andrea Braun-Kiewnick, Jan Tedrup, Carla Meijerink, Elena Durer, Claudia Ritter, Peter M Keller, Michael Hombach
Objectives: This study evaluated the AID carbapenemase line probe assay (LPA) for the detection and identification of carbapenem resistance genes in Enterobacteriaceae and other Gram-negative bacilli (GNB) using bacterial cultures and DNA extracts directly from patient urine samples. Methods: The AID carbapenemase LPA detects 13 different carbapenemase genes. Test probe accuracy was verified for using clinical Enterobacteriaceae isolates harbouring bla KPC , bla VIM , bla NDM , bla GIM , bla AIM , bla SPM , bla IMP and bla OXA-48 and a well-characterized set of Escherichia coli DH5α strains transformed with the vector plasmid pUC57- kan harbouring bla BIC , bla SIM , bla DIM , bla IMI-3 , bla IMI-1 and bla NMC-A ...
April 11, 2017: Journal of Antimicrobial Chemotherapy
https://www.readbyqxmd.com/read/28401351/quantification-of-intrahepatic-total-hbv-dna-in-liver-biopsies-of-hbv-infected-patients-by-a-modified-version-of-cobas-%C3%A2-ampliprep-cobas-%C3%A2-taqman-hbv-test-v2-0
#14
Romina Salpini, Lorenzo Piermatteo, Upkar Gill, Arianna Battisti, Francesca Stazi, Tania Guenci, Sara Giannella, Valentina Serafini, Patrick T F Kennedy, Carlo Federico Perno, Valentina Svicher, Marco Ciotti
Intrahepatic total HBV DNA (it-HBV DNA) level might reflect the size of virus reservoir and correlate with the histological status of the liver. To quantitate it-HBV DNA in a series of 70 liver biopsies obtained from hepatitis B chronic patients, a modified version of the COBAS(®)Ampliprep/COBAS(®)TaqMan HBV test v2.0 was used for this purpose. The linearity and reproducibility of the modified protocol was tested by quantifying serial dilutions of a full-length HBV containing plasmid and it-HBV DNA from a reference patient...
April 11, 2017: Medical Microbiology and Immunology
https://www.readbyqxmd.com/read/28389898/influence-of-sucrose-on-growth-and-sensitivity-of-candida-albicans-alone-and-in-combination-with-enterococcus-faecalis-and-streptococcus-mutans-to-photodynamic-therapy
#15
Fernanda Malagutti Tomé, Lucas De Paula Ramos, Fernanda Freire, Cristiane Aparecida Pereira, Ingrid Christine Barbosa de Oliveira, Juliana Campos Junqueira, Antonio Olavo Cardoso Jorge, Luciane Dias de Oliveira
This study has evaluated the effects of photodynamic inactivation (PDI) using erythrosine as photosensitizer and green light-emitting diode (LED) on biofilms of Candida albicans alone and in combination with Enterococcus faecalis and Streptococcus mutans. We have also evaluated the effect of sucrose on biofilm formation and bacterial growth and sensitivity to PDI. Biofilms were formed in suspension of 10(6) cells/ml on plates before being grown in broth culture with and without sucrose and incubated for 48 h...
April 7, 2017: Lasers in Medical Science
https://www.readbyqxmd.com/read/28388918/tea-camellia-sinensis-infusions-ameliorate-cancer-in-4ti-metastatic-breast-cancer-model
#16
Karori S Mbuthia, Paul O Mireji, Raphael M Ngure, Francesca Stomeo, Martina Kyallo, Chalo Muoki, Francis N Wachira
BACKGROUND: Tea (Camellia sinensis) infusions are widely consumed beverages with numerous health benefits. However, physiological and molecular responses mediating these activities are poorly understood. METHOD: Three replicates of 4TI cancer cell suspension (2.0 × 10(5) cells/ml) were challenged in vitro with various concentrations of green, black and purple tea infusions to asseses their cytoxicity and associated differentially expressed genes in the cells...
April 7, 2017: BMC Complementary and Alternative Medicine
https://www.readbyqxmd.com/read/28381602/cerebrospinal-fluid-treponema-pallidum-particle-agglutination-assay-for-neurosyphilis-diagnosis
#17
Christina M Marra, Clare L Maxwell, Shelia B Dunaway, Sharon K Sahi, Lauren C Tantalo
BACKGROUND: Limited data suggest that the cerebrospinal fluid (CSF) Treponema pallidum particle agglutination assay (TPPA) is sensitive and a CSF Treponema pallidum hemagglutination assay (TPHA) titer ≥1:640 is specific for neurosyphilis diagnosis. METHODS: CSF-TPPA reactivity and titer were determined for a convenience sample of 191 CSFs from individuals enrolled in a study of CSF abnormalities in syphilis (training dataset). Sensitivity of a reactive test and specificity for reactivity at serial higher CSF dilutions was determined...
April 5, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28375717/optimization-of-a-plaque-neutralization-test-pnt-to-identify-the-exposure-history-of-pacific-herring-to-viral-hemorrhagic-septicemia-virus-vhsv
#18
Lucas M Hart, Ashley MacKenzie, Maureen K Purcell, Rachel L Powers, Paul K Hershberger
Methods for a plaque neutralization test (PNT) were optimized for the detection and quantification of viral hemorrhagic septicemia virus (VHSV) neutralizing activity in the plasma of Pacific Herring Clupea pallasii. The PNT was complement dependent, as neutralizing activity was attenuated by heat inactivation; further, neutralizing activity was mostly restored by the addition of exogenous complement from specific-pathogen-free Pacific Herring. Optimal methods included the overnight incubation of VHSV aliquots in serial dilutions (starting at 1:16) of whole test plasma containing endogenous complement...
June 2017: Journal of Aquatic Animal Health
https://www.readbyqxmd.com/read/28372840/restriction-capillaries-as-an-innovative-mixing-unit-for-intermediate-mobile-phase-exchange-in-multidimensional-analysis
#19
Glenn Loos, Konstantin Shoykhet, Monika Dittmann, Deirdre Cabooter
A novel mixing unit is proposed for the serial coupling of orthogonal columns to analyze polar and non-polar compounds in a single run. The principle relies on the isolation of unretained peaks eluting from a first dimension column in a sample loop, before directing them to a second column for separation. Since the mobile phases employed in highly orthogonal separations are not directly compatible, a mixing unit is required to alter the mobile phase composition before executing the second dimension separation...
March 19, 2017: Journal of Chromatography. A
https://www.readbyqxmd.com/read/28370080/evaluation-of-assays-for-quantification-of-dna-in-canine-plasma-as-an-indirect-marker-of-netosis
#20
Stephanie A Smith, Corinne M Lawson, Maureen A McMichael, Katrina Jung, Mauria O'Brien, Ron Achiel
BACKGROUND: Neutrophil extracellular traps (NET), consisting of a filamentous DNA/chromatin-histone scaffold originating from neutrophils are part of the innate immune response, may be released under a variety of inflammatory conditions and are associated with an increased risk for thrombosis. OBJECTIVES: The purpose of this study was to evaluate a SYTOX green fluorescence assay and a histone-DNA complex (hisDNA) ELISA for quantification of NET-related DNA in canine plasma...
April 3, 2017: Veterinary Clinical Pathology
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