Name: production of high-titer infectious influenza pseudotyped particles with envelope glycoproteins from highly pathogenic h5n1 and avian h7n9 viruses
Uploaded: 2020-01-15T12:00:00
Description: Watch this Scientific Journal Video about Production of High-Titer Infectious Influenza Pseudotyped Particles with Envelope Glycoproteins from Highly Pathogenic H5N1 and Avian H7N9 Viruses at JoVE.com

Dette arbejde blev støttet af tilskud fra Zhejiang Provincial medicin og sundhed videnskab og teknologiplan (Grant numre, 2017KY538), Hangzhou kommunale medicin og sundhed videnskab og teknologiplan (Grant numre, OO20190070), Hangzhou Medical Science og Teknologi nøgleprojekt (tilskuds numre, 2014Z11) og Hangzhou Municipal selvstændigt applikations projekt for social udvikling og videnskabelig forskning (tilskuds numre, 20191203B134).

1. dag 1: cellekultur og såning
<ol><li>Dyrke menneskelige embryonale nyrer (HEK) 293T/17 celler i 60 mm retter med Dulbecco's modificerede essentielle medium (DMEM) suppleret med 10% føtal bovin serum (FBS) og 100 U/mL penicillin-streptomycin (DMEM komplet medium, DCM) i en 37 °C, 5% carbondioxid (CO2) inkubator indtil omkring 80% konflydende. Bemærk: HEK 293T/17 Low passage celler anbefales.</li>
	<li>Cellerne vaskes forsigtigt med 5 mL fosfat bufferet saltvand (PBS) 1x. Bemærk: manuel håndteri...

<ol>
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I denne protokol beskriver vi en metode til fremstilling af pseudo-viruspartikler (PP) i en BSL-2-indstilling. Indberetteren plasmid pcDNA-GFP er indarbejdet i PPS og kan anvendes til at kvantificere KKS ved FACER i en infektivitetsanalyse. Vi valgte to typer af modtagelige cellelinjer, fordi de er meget udbredt i influenza forskning. Mdck celler ville give en god kontrol til de variable udødeliggjort humane celler, der anvendes i disse undersøgelser.
Denne protokol er baseret på retrovirus...

Missionen af en viral partikel er at transportere sit genom fra en inficeret værtscelle til en ikke-inficeret værtscelle og at levere den til cytoplasmaet eller kernen i en replikeringskompetent formular1. Denne proces er oprindeligt udløst af binding til Host celle receptorer, efterfulgt af fusion af virion og cellulære membraner. For kappeklædte vira, som influenzavirus, er Spike glycoproteiner ansvarlig for receptor binding og fusion1,2. Viral konvolut glycoproteiner (f. eks. pyrogener, antigener), er involveret i mange vigtige egenskaber og begivenheder, såsom virus livscyklus initiering (binding og fusion), viral patogenese, immunogenicitet, Host Cell apoptose og cellulære tropisme, den cellulære endocytic pathway, samt interspecies transmission og resortiments1,3,4,5,6,7. Forskning på viral konvolut glycoproteiner vil hjælpe os med at forstå mange aspekter af den virale infektions proces. Pseudo-skrevne virale partikler (PP), også kaldet pseudovirioner eller pseudopartikler, kan genereres gennem en pseudotyping Technique8,9,10. Denne teknologi er blevet brugt til at udvikle pseudo-skrevne partikler af mange vira, herunder hepatitis C11,12, hepatitis B13, vesikulær stomatitis virus (VSV)14,15og influenzavirus16,17,18,19. Denne teknologi er baseret på gag-pol protein af lentivira eller andre retrovira.
Pseudoskrevne virale partikler kan opnås ved hjælp af et tre-plasmid-system ved at cotransfficerer en viral konvolut glykoprotein ekspression plasmid, en antiretroviral emballage plasmid mangler konvolutten env genet, og en separat reporter plasmid i PP producer celler. Retrovirus er samlet af dens gag protein, og det knopper fra en inficeret celle membran, der udtrykker virus konvolut protein1. Derfor er det muligt at opnå høj titer influenza PPS ved hjælp af retrovirus gag protein til at producere knopper på en cellulær membran, der udtrykker influenza HA og NA. I vores tidligere undersøgelser, har/NAS i alle kombinationer var funktionelle og i stand til at udføre deres tilsvarende funktioner i viral livscyklus16,17,18,20,21. Disse KKS anvendes til at undersøge influenza biologiske egenskaber, herunder hemagglutination, neuraminidaseaktivitet, HA-receptor bindende tropisme og infektivitet. Fordi HA og NA begge er vigtige overflade funktionelle proteiner i den virale livscyklus, har uoverensstemmende og NAs afledt af forskellige stammer af influenza delvist kan demonstrere resortiments mellem dem. Her genererer vi otte typer af influenza PPS ved at kombinere to har og to NAs (afledt af HPAI H5N1 stamme og H7N9 Stain), ved hjælp af en tre-plasmid pseudotyping system. Disse otte typer PPS omfatter to Native PPS, H5N1pp, H7N9pp; to uoverensstemmende KKS (H5 + N9) PP, (H7 + N1) PP; og fire PPS kun harkedeligt en enkelt glykoprotein (ha eller na), H5pp, N1pp, H7pp, N9pp. undersøgelser af influenzavirus, såsom H5N1 og H7N9, er begrænset af biosikkerhedskrav. Alle undersøgelser af vilde influenzavirusstammer bør udføres på et biosikkerhedsniveau 3 (BSL-3) laboratorium. Pseudotyp viral partikel teknologi kan bruges til at pakke en kunstig virion i en biosikkerhedsniveau 2 (BSL-2) indstilling. Derfor er PPS et sikrere og nyttigt redskab til at studere influenzavirus processer afhængigt af dens to store glycoproteiner: hemagglutinin (HA) og neuraminidasetypen (Na).
Denne protokol beskriver genereringen af disse PPS med en tre-plasmid cotransfection strategi (oversete i figur 1), hvordan man kvantificerer PPS, og infektivitet påvisning. PP-produktionen involverer tre typer plasmid'er (figur 1). Gag-pol genet, som koder retrovirus gag-pol protein, blev klonet fra en retrovirus emballage kit og indsat i pcdna 3,1 plasmid og hedder Pcdna-gag-pol. Det forbedrede grønne fluorescerende protein (eGFP)-gen, som koder grønt fluorescerende protein, blev klonet fra pTRE-EGFP-vektor, indsat i pcDNA 3,1 plasmid, og kaldet pcDNA-GFP. Under kloning blev en pakke signal sekvens tilføjet via en primer. HA-og NA-gener blev klonet ind i en pVRC-plasmid, der blev henholdsvis pVRC-HA og pVRC-NA. Det sidste plasmid koder fusions proteinet og kan udskiftes med ethvert andet fusionsprotein af interesse. Vores pseudo Typing platform indeholder to glykoprotein udtryks plasmider: pvrc-ha og pvrc-na. Dette kan forenkle forskningen i omsortimentet mellem forskellige virusstammer i en BSL-2-indstilling.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Benzonase Nuclease</td>
			<td>Millipore</td>
			<td>70664</td>
			<td>Effective viscosity reduction and removal of nucleic acids from protein solutions</td>
		</tr>
		<tr>
			<td>Clear Flat Bottom Polystyrene TC-treated Microplates (96-well)</td>
			<td>Corning</td>
			<td>3599</td>
			<td>Treated for optimal cell attachment 
			Sterilized by gamma radiation and certified nonpyrogenic 
			Individual alphanumeric codes for well identification</td>
		</tr>
		<tr>
			<td>Clear TC-treated Multiple Well Plates (6-wells)</td>
			<td>Costar</td>
			<td>3516</td>
			<td>Individual alphanumerical codes for well identification 
			Treated for optimal cell attachment 
			Sterilized by gamma irradiation</td>
		</tr>
		<tr>
			<td>Dulbecco&#39;s modified essential medium (DMEM)</td>
			<td>Gibco</td>
			<td>11965092</td>
			<td>A widely used basal medium for supporting the growth of many different mammalian cells</td>
		</tr>
		<tr>
			<td>Fetal bovine serum</td>
			<td>Excell</td>
			<td>FND500</td>
			<td>fetal bovine sera that can offer excellent value for basic cell culture, specialty research, and specific assays</td>
		</tr>
		<tr>
			<td>Fluorescence Activated Cell Sorting (FACS)</td>
			<td>Beckman coulter</td>
			<td>cytoflex</td>
			<td></td>
		</tr>
		<tr>
			<td>Human alveolar adenocarcinoma A549 cells</td>
			<td>ATCC</td>
			<td>CRM-CCL-185</td>
			<td></td>
		</tr>
		<tr>
			<td>Human embryonic kidney (HEK) HEK-293T/17 cells</td>
			<td>ATCC</td>
			<td>CRL-11268</td>
			<td>A versatile transfection reagent that has been shown to effectively transfect the widest variety of adherent and suspension cell lines</td>
		</tr>
		<tr>
			<td>Inverted fluorescent biological microscope</td>
			<td>Olympus</td>
			<td>BX51-32P01-FLB3</td>
			<td></td>
		</tr>
		<tr>
			<td>Inverted light microscope</td>
			<td>Olympus</td>
			<td>CKX31-12PHP</td>
			<td></td>
		</tr>
		<tr>
			<td>Lipofectamine 2000 Transfection Reagent</td>
			<td>Invitrogen</td>
			<td>11668019</td>
			<td>Rapid, sensitive and precise probe-based qPCR detection and quantitation of target RNA targets.</td>
		</tr>
		<tr>
			<td>Luna Universal Probe One-Step RT-qPCR Kit</td>
			<td>NEB</td>
			<td>E3006L</td>
			<td>Will withstand up to 14,000 RCF 
			RNase-/DNase-free Nonpyrogenic</td>
		</tr>
		<tr>
			<td>Madin-Darby Canine Kidney (MDCK) cells</td>
			<td>ATCC</td>
			<td>CCL-34</td>
			<td></td>
		</tr>
		<tr>
			<td>MaxyClear Snaplock Microcentrifuge Tube (1.5 mL)</td>
			<td>Axygen</td>
			<td>MCT-150-C</td>
			<td>33 mm, gamma sterilized</td>
		</tr>
		<tr>
			<td>Millex-HV Syringe Filter Unit, 0.45 &#181;m, PVDF</td>
			<td>Millipore</td>
			<td>SLHV033RS</td>
			<td>an improved Minimal Essential Medium (MEM) that allows for a reduction of Fetal Bovine Serum supplementation by at least 50% with no change in cell growth rate or morphology. Opti-MEM I medium is also recommended for use with cationic lipid transfection reagents, such as Lipofectamine reagent.</td>
		</tr>
		<tr>
			<td>Opti-MEM I Reduced Serum Medium</td>
			<td>Gibco</td>
			<td>11058021</td>
			<td>The antibiotics penicillin and streptomycin are used to prevent bacterial contamination of cell cultures due to their effective combined action against gram-positive and gram-negative bacteria.</td>
		</tr>
		<tr>
			<td>penicillin-streptomycin</td>
			<td>Gibco</td>
			<td>15140122</td>
			<td>Maximum RCF is 12,500 xg 
			Temperature range from -80 &#176;C to 120 &#176;C 
			RNase-/DNase-free 
			Sterile</td>
		</tr>
		<tr>
			<td>PP Centrifuge Tubes (15 mL)</td>
			<td>Corning</td>
			<td>430791</td>
			<td>a stable and highly reactive serine protease</td>
		</tr>
		<tr>
			<td>Proteinase K</td>
			<td>Beyotime</td>
			<td>ST532</td>
			<td>Treated for optimal cell attachment 
			Sterilized by gamma radiation and certified nonpyrogenic</td>
		</tr>
		<tr>
			<td>TC-treated Culture Dish (60mm)</td>
			<td>Corning</td>
			<td>430166</td>
			<td>Trypsin from bovine pancreas 
			TPCK Treated, essentially salt-free, lyophilized powder, &#8805;10,000 BAEE units/mg protein</td>
		</tr>
		<tr>
			<td>TPCK-trypsin</td>
			<td>Sigma</td>
			<td>T1426</td>
			<td>This liquid formulation of trypsin contains EDTA and phenol red. Gibco Trypsin-EDTA is made from trypsin powder, an irradiated mixture of proteases derived from porcine pancreas. Due to its digestive strength, trypsin is widely used for cell dissociation, routine cell culture passaging, and primary tissue dissociation. The trypsin concentration required for dissociation varies with cell type and experimental requirements.</td>
		</tr>
		<tr>
			<td>Trypsin-EDTA (0.25%), phenol red</td>
			<td>Gibco</td>
			<td>25200056</td>
			<td></td>
		</tr>
	</tbody>
</table>

production of high-titer infectious influenza pseudotyped particles with envelope glycoproteins from highly pathogenic h5n1 and avian h7n9 viruses

Lejlighedsvis direkte overførsel af højpatogen aviær influenza A-virus H5N1 (HPAI H5N1) og H7N9 til mennesker og deres dødelighed er alvorlige folkesundhedsproblemer og antyder muligheden for en epidemi. Men vores molekylære forståelse af virussen er rudimentær, og det er nødvendigt at studere de biologiske egenskaber af dens konvolut proteiner som terapeutiske mål og udvikle strategier til at kontrollere infektion. Vi udviklede en solid viral pseudo-type partikel (PP)-platform til at studere aviær influenzavirus, herunder den funktionelle analyse af dens hemagglutinin (HA) og neuraminidasetypen (Na) konvolut glycoproteiner, resortiments egenskaberne af HAs og NAs, receptorer, tropismer, neutraliserende antistoffer, diagnose, infektivitet, med henblik på narkotika udvikling og vaccine design. Her beskriver vi en eksperimentel procedure for at etablere PPS med kuvert glycoproteiner (HA, NA) fra to influenza A-stammer (HAPI H5N1 og 2013 aviær H7N9). Deres generation er baseret på kapaciteten af nogle vira, såsom murine leukæmivirus (MLV), at indarbejde kuvert glycoproteiner i en PP. Desuden er vi også detaljeret, hvordan disse PPS er kvantificeret med RT-qPCR, og infektivitet påvisning af indfødte og uoverensstemmende virus PPS afhængigt af oprindelsen af HAs og NAs. Dette system er meget fleksibelt og tilpasningsdygtigt og kan bruges til at etablere virale PPS med kuvert glycoproteiner, der kan inkorporeres i enhver anden form for kappeklædte virus. Således kan denne viral partikel platform bruges til at studere vilde vira i mange forskningsundersøgelser.

Afhængigt af den generelle procedure, der er beskrevet ovenfor, har vi genereret 10 typer af PPS kombinere to gruppe har/NAS eller VSV-G glykoprotein eller no-konvolut glycoproteiner (vist i tabel 1). Syv af dem er smitsomme. PPS, at havnen No-konvolut glykoprotein eller kun havn na ikke vise nogen infektivitet her. Produktions proceduren for influenza-PP er oversete i figur 1. Transmissions elektron mikrografer af PPS (f. eks. H5N1pp) er vist i figur 3...

Watch this Scientific Journal Video about Production of High-Titer Infectious Influenza Pseudotyped Particles with Envelope Glycoproteins from Highly Pathogenic H5N1 and Avian H7N9 Viruses at JoVE.com

Production of High-Titer Infectious Influenza Pseudotyped Particles with Envelope Glycoproteins from Highly Pathogenic H5N1 and Avian H7N9 Viruses

Denne protokol beskriver en eksperimentel proces til fremstilling af infektiøse virale pseudo skrevne partikler (PP) med kuvert glycoproteiner fra to influenza A-stammer, og hvordan deres infektionsevne kan bestemmes. Denne protokol er meget tilpasningsdygtig til at udvikle PPS af enhver anden type kappeklædte vira med forskellige konvolut glycoproteiner.

Produktion af Højtiter infektiøse influenza Pseudoskrevne partikler med kuvert glycoproteiner fra højpatogen H5N1 og aviær H7N9 vira

The occasional direct transmission of the highly pathogenic avian influenza A virus H5N1 (HPAI H5N1) and H7N9 to humans and their lethality are serious ...

Denne teknik til fremstilling af influenza viral PP og bestemmelse af dens infektivitet kan forenkle forskningen i influenzavirus i en BSL-2 by. Vi normaliserede PPS for infektioner baseret på qRT-PCR data af PPS. To glycoprotein udtryk plasmid er kodet, hvilket kan forenkle forskningen på virus reassortment.En dag efter cellesåning kontrolleres cellemorfologien og tætheden under et omvendt lysmikroskop. Ideelt set cellen skal være ca 85% sammenløb ved transfektion. Udskift mediet med en milliliter serumfri DMEM pr. brønd, og læg pladen tilbage i inkubatoren.For hver brønd af celler, der skal transsmittede, fortyndes otte mikroliter af transfektionsreageret til et volumen på 150 mikroliter med reduceret serummedium. Opløsningen blandes forsigtigt, og den skal sidde ved stuetemperatur i fem minutter. I mellemtiden fortyndes 2,5 mikrogram plasmid DNA i 158 mikroliter af reduceret serummedium.Efter fem minutters inkubation kombineres det fortyndede DNA med fortyndet transinfektionsreage, opløsningen blandes forsigtigt og den efterlades ved stuetemperatur i 15 minutter. Tilsæt DNA lipidkomplekset til de tilsvarende brønde med cellerne i serumfrit medium. Bland derefter pladen forsigtigt ved at rokke frem og tilbage.Inkuber pladen ved 37 grader Celsius og 5% kuldioxid i fire til seks timer. Efter inkubationen fjernes mediet og udskiftes med to milliliter DMEM pr. brønd. Derefter inkubere det i yderligere 36 til 48 timer.Frø hver type modtagelig celle på 10, 000 celler pr brønd og en 96 brønd plade. Derefter forlade pladen i løbet af natten i en 37 grader Celsius og fem procent kuldioxid inkubator. På den tredje dag efter transfektion, kontrollere farven på mediet, som skal lyse pink eller lidt orange og undersøge cellerne med en omvendt lysstofsnit biologisk mikroskop under 440 til 460 nanometer bølgelængde.På 38 til 48 timer efter efter transfektion, høste pseuotyped partikler eller PPS ved at passere dem gennem en 0,45 mikrometer polyvinylidene fluormembranfilter for at fjerne celleaffald, derefter opdele dem i små volumen aliquots og gemme dem på minus 80 grader Celsius. For at kvantificere PPS overføres 30 mikroliter af de rensede viruspartikler til et 1,5 mikroliter RNase-frit rør, og der tilsættes en mikroliter Benzonase Nuclease. Inkuber røret ved 37 grader Celsius i en time for at eliminere enhver DNA- og RNA-forurening.Efter inkubationen fryses prøven til minus 70 grader Celsius for at aktivgøre nuklease og tilsættes to mikroliter proteinase K.Inkuber blandingen i 30 minutter for at fordøje konvolutproteinerne og frigive CMV-GFP RNA. Derefter inaktive Proteinase K ved 100 grader Celsius i tre minutter. Kvantificer PPS med kvantitativ omvendt transskription RTPCR ved hjælp af Universal Probe One-Step RT-qPCR Kit i primere og sonder, der er angivet i tekstmanuskriptet.Normalisere PPS til fire gange 10 til femte RNA kopier pr. milliliter. Derefter tilsættes TPCK-trypsin til en endelig koncentration på 10 mikrogram pr. milliliter i PPS, der huser H7N9 hemagglutinin. Inkuber KKS ved 37 grader Celsius i en time for at danne de funktionelle underenheder HA1 og HA2.Den normaliserede PPS med DMEM-mediet blandes i et forhold på 1-1, hvortil derefter pladen, der indeholder modtagelige celler, til biosikkerhedsskabet. Inspirer supernatanten og vask cellerne én gang med 100 mikroliter af forvarmt PBS. Der tilsættes 100 mikroliter PPS-DMEM-blanding i hver brønd, hvilket giver infektivitetstest af hver type PPS til en modtagelig cellelinje.Inkuber pladen i fire til seks timer ved 37 grader Celsius og 5% kuldioxid. Derefter aspirere supernatant og erstatte det med 100 mikroliter af DCM Inkubere pladen i yderligere 24 til 36 timer før infektivitet afsløring. Vores resultater er PPS anses nu smitsom for mennesker.Så nødvendige sikker vagt procedurer anbefales at tage såsom at bære en maske og udføre infektivitet assays i en biosikkerhed kabinet. Denne protokol blev genereret til at generere 10 typer af pseudotypede partikler eller PPS med to grupperet hemaglutinin og neuraminidase vesikulær stomatitis virus G glycoprotein eller ingen kuvert glycoproteiner. PPS blev afbildet med transmission elektronmikroskopi og deres infektivitet blev evalueret i to cellelinjer, A549 og MDCK.I PPS-gruppen, der huser H5, var infektiviteten af H5N1, H5 plus N9 og H5 henholdsvis ca. 90%18% og 10% for cellelinje A549 og 40%5% og 5% for MDCK. I PPS-koncernen, der huser H7, var smitteevnen for H7N9, H7 plus N1 og H7 henholdsvis ca. 10%7% og 1% for cellelinje A549 og 8%4% og 1% for MDCK. Infektiviteten af vesikulær stomatitis virus G glycoprotein PP var omkring 21% for A549 og 16% for MDCK celler.Mens delta kuvert glycoproteiner PP viste ingen smitteevne. Det er vigtigt at huske, at PP producent HEK-293T/17 celler udgør grundlaget for denne procedure. Så den optimale vækst lærer os HEK-293T/17 celler er vigtigst.Efter denne procedure eller en anden test såsom L2 recept afgrænser assay kan udføres. Denne analyse kan næsten give slippe af med de biologiske særheder af receptor præference og mønsteret vil afsløre tropisme af PPS. Da de tog glycoproteiner fra influenzavirus er klonet i to plus max.HA og NA proteiner kan studeres separat, så vi reassortment mellem dem. Mutanterne og de modne effekter kan udforskes.

Research

JoVE Journal

Immunology and Infection

Determination of Molecular Structures of HIV Envelope Glycoproteins using Cryo-Electron Tomography and Automated Sub-tomogram Averaging

Bestemmelse af molekylære strukturer af HIV Konvolut glycoproteiner med Cryo-Electron Tomography og Automatiseret Sub-tomogram Gennemsnitsberegnings

Since its discovery nearly 30 years ago, more than 60 million people have been infected with the human immunodeficiency virus (HIV) (www.usaid.gov). The ...

Avian Influenza Surveillance with FTA  Cards: Field Methods, Biosafety, and Transportation Issues Solved

Avian Influenza Viruses (AIVs) infect many mammals, including humans1. These AIVs are diverse in their natural hosts, harboring almost all possible viral ...

Packaging HIV- or FIV-based Lentivector Expression Constructs &amp; Transduction of VSV-G Pseudotyped Viral Particles

Packaging HIV- or FIV-based Lentivector Expression Constructs & Transduction of VSV-G Pseudotyped Viral Particles

Emballagen HIV-eller FIV-baserede Lentivector ekspressionskonstruktioner og transduktion af VSV-G-pseudotypet virale partikler

As with standard plasmid vectors, it is possible to transfect lentivectors in plasmid form into cells with low-to-medium efficiency to obtain transient ...

Expression of Functional Recombinant Hemagglutinin and Neuraminidase Proteins from the Novel H7N9 Influenza Virus Using the Baculovirus Expression System

Ekspression af funktionelle Rekombinant hæmagglutinin og neuraminidase Proteiner fra Novel H7N9 influenza virus ved hjælp baculovirusekspressionssystemet

The baculovirus expression system is a powerful tool for expression of  recombinant proteins. Here we use it to produce correctly folded and  glycosylated ...

Conformational Evaluation of HIV-1 Trimeric Envelope Glycoproteins Using a Cell-based ELISA Assay

Konformationel Evaluering af HIV-1 trimere kappeglycoproteinerne hjælp af en celle-ELISA-assay

HIV-1 envelope glycoproteins (Env) mediate viral entry into target cells and are essential to the infectious cycle. Understanding how those glycoproteins ...

Modeling The Lifecycle Of Ebola Virus Under Biosafety Level 2 Conditions With Virus-like Particles Containing Tetracistronic Minigenomes

Modeling Lifecycle af Ebola virus Under biosikkerhedsniveau 2 Betingelser med virus-lignende partikler indeholdende Tetracistronic minigenomer

Ebola viruses cause severe hemorrhagic fevers in humans and non-human primates, with case fatality rates as high as 90%. There are no approved vaccines or ...

Intranasal Administration of Recombinant Influenza Vaccines in Chimeric Mouse Models to Study Mucosal Immunity

Intranasal administration af rekombinant influenzavacciner i kimære musemodeller at studere slimhindeimmunitet

Vaccines are one of the greatest achievements of mankind, and have saved millions of lives over the last century. Paradoxically, little is known about the ...

Treatment of Platelet Products with Riboflavin and UV Light: Effectiveness Against High Titer Bacterial Contamination

Behandling af blodplade Produkter med riboflavin og UV-lys: effektivitet mod højtiter bakteriel forurening

Contamination of platelet units by bacteria has long been acknowledged as a significant transfusion risk due to their post-donation storage conditions. ...

Rapid Molecular Detection and Differentiation of Influenza Viruses A and B

Hurtig Molekylær Detection og Differentiering af Influenza Virus A og B

Influenza is a contagious respiratory illness caused by influenza viruses A and B in humans and causes a significant amount of morbidity and mortality ...

Fluorescence-based Neuraminidase Inhibition Assay to Assess the Susceptibility of Influenza Viruses to The Neuraminidase Inhibitor Class of Antivirals

Fluorescensbaseret neuraminidase-inhiberingsassay i vurdering af den følsomhed over influenzavira til neuraminidaseinhibitoren Class of Antivirale

The neuraminidase (NA) inhibitors are the only class of antivirals approved for the treatment and prophylaxis of influenza that are effective against ...