Name: a high-throughput platform for the screening of salmonella spp./shigella spp.
Uploaded: 2018-11-07T14:00:00
Description: Watch this Scientific Journal Video about A High-throughput Platform for the Screening of Salmonella spp./Shigella spp. at JoVE.com

Dette arbejde blev støttet af videnskab og teknologi Program af Zhuhai, Kina (grant nummer 20171009E030064), videnskab og teknologi Program af Guangdong, Kina (grant nummer 2015A020211004) og videnskab og teknologi Program af General Administration for kvalitetsovervågning, inspektion og karantæne i Folkerepublikken Kina (grant nummer 2016IK302, 2017IK224).

Protokollen følger retningslinjerne, af den menneskelige videnskabsetisk Komité i Zhuhai International Travel Healthcare Center. Brug venligst standard sterile operation under eksperimentet.
1. kultur medier sammensætning og forberedelse
<ol><li>Forbereder næringsstoffer bouillon: Opløs 1% pepton, 0,3% oksekød ekstrakt, 0,5% natriumchlorid og 0,1% glukose i H2O, justere pH 7,5 og autoklave det i 121 ° C i 15 min.</li>
	<li>Forberede Selenite cystin medium: Opløs 0,5% pepto...

<ol>
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(258), 89-92 (2016).</li><li>Li, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Establishment+and+Application+of+a+Visual+DNA+Microarray+for+the+Detection+of+Food-borne+Pathogens.">Establishment and Application of a Visual DNA Microarray for the Detection of Food-borne Pathogens.</a> Analytical Sciences. 32 (2), 215-218 (2016).</li><li>Zhuang, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+Salmonella+spp.+by+a+loop-mediated+isothermal+amplification+(LAMP)+method+targeting+bcfD+gene.">Detection of Salmonella spp. by a loop-mediated isothermal amplification (LAMP) method targeting bcfD gene.</a> Letters in Applied Microbiology. 59 (6), 658-664 (2014).</li><li>Shi, X. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+simultaneous+detection+of+Salmonella+and+Shigella+using+modified+molecular+beacons+and+real-time+PCR.">Rapid simultaneous detection of Salmonella and Shigella using modified molecular beacons and real-time PCR.</a> Zhonghua Liu Xing Bing Xue Za Zhi. 27 (12), 1053-1056 (2006).</li><li>Mo, Q. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+of+a+96-microwell+plate+DNA+diagnostic+chip+for+detection+of+foodborne+bacteria+and+its+application+in+an+incident+of+food+poisoning.">Preparation of a 96-microwell plate DNA diagnostic chip for detection of foodborne bacteria and its application in an incident of food poisoning.</a> Nan Fang Yi Ke Da Xue Xue Bao. 30 (3), 417-421 (2010).</li><li>Wang, H. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Probe-free+and+sensitive+detection+of+diarrhea-causing+pathogens+using+RT-PCR+combined+high+resolution+melting+analysis.">Probe-free and sensitive detection of diarrhea-causing pathogens using RT-PCR combined high resolution melting analysis.</a> Biologicals. 44 (5), 360-366 (2016).</li><li>Sun, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+simultaneous+screening+of+seven+clinically+important+enteric+pathogens+using+a+magnetic+bead+based+DNA+microarray.">Rapid simultaneous screening of seven clinically important enteric pathogens using a magnetic bead based DNA microarray.</a> World Journal of Microbiology and Biotechnology. 27 (1), 163-169 (2011).</li><li>Qi, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiplex+PCR+assay+for+rapid+detection+of+five+important+pathogenic+vibrios.">Multiplex PCR assay for rapid detection of five important pathogenic vibrios.</a> Chinese Journal of health laboratory technology. (24), 3497-3500 (2014).</li><li>Blanco, G., Diaz de Tuesta, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Culture-+and+molecular-based+detection+of+swine-adapted+Salmonella+shed+by+avian+scavengers.">Culture- and molecular-based detection of swine-adapted Salmonella shed by avian scavengers.</a> Science of the Total Environment. 634, 1513-1518 (2018).</li><li>Tang, X. J., Yang, Z., Chen, X. B., Tian, W. F., Tu, C. N., Wang, H. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Verification+and+large+scale+clinical+evaluation+of+a+national+standard+protocol+for+Salmonella.spp./Shigella.spp.+screening+using+real-time+PCR+combined+with+guided+culture.">Verification and large scale clinical evaluation of a national standard protocol for Salmonella.spp./Shigella.spp. screening using real-time PCR combined with guided culture.</a> Journal of Microbiological Methods. 145, 14-19 (2018).</li><li>Dekker, D. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drinking+water+from+dug+wells+in+rural+ghana--salmonella+contamination,+environmental+factors,+and+genotypes.">Drinking water from dug wells in rural ghana--salmonella contamination, environmental factors, and genotypes.</a> International Journal of Environmental Research and Public Health. 12 (4), 3535-3546 (2015).</li><li>Gargano, J. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mortality+from+selected+diseases+that+can+be+transmitted+by+water+-+United+States,+2003-2009.">Mortality from selected diseases that can be transmitted by water - United States, 2003-2009.</a> Journal of Water and Health. 15 (3), 438-450 (2017).</li><li>Kumar, R., Surendran, P. K., Thampuran, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+culture,+ELISA+and+PCR+assays+for+the+detection+of+Salmonella+in+seafood.">Evaluation of culture, ELISA and PCR assays for the detection of Salmonella in seafood.</a> Letters in Applied Microbiology. 46 (2), 221-226 (2008).</li><li>Herrera-Leon, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blind+comparison+of+traditional+serotyping+with+three+multiplex+PCRs+for+the+identification+of+Salmonella+serotypes.">Blind comparison of traditional serotyping with three multiplex PCRs for the identification of Salmonella serotypes.</a> Research in Microbiology. 158 (2), 122-127 (2007).</li><li>Cunningham, S. 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Siden Salmonella spp. /Shigella spp. er ofte forbundet med madforgiftning og fækal-oral transmission af akut gastroenteritis28,29 og rutinemetoden er enten arbejdskrævende eller tidskrævende 7, vi beskriver en høj overførselshastighed platform for Salmonella spp. /Shigella spp. screening, ved hjælp af real-time PCR kombineret med guidet kultur.
Der er flere trin, der har...

Diarré er stadig en fælles spørgsmål om sundhed med en høj incidens Vurder globalt1,2. Selvom dødeligheden er relativt lav, nogle patienter viser forskellige symptomer for uger (f.eks. løs og vandig afføring, en hastesag at gå på toilettet), hvilket gør den socioøkonomiske indvirkning meget høj3,4. Mere alvorligt, nogle patienter kan selv udvikle irritabel tyktarm, hvis venstre ubehandlet5. Der findes mange forskellige slags bakterier, virus og parasitter, der kan forårsage diarré6. Salmonella spp. /Shigella spp. er blandt de mest almindelige bakterier til transmission af akut gastroenteritis7,8,9,10,11. Derfor, mange amter har udstedt love eller regler for regelmæssig Salmonella spp. /Shigella spp. screening blandt mennesker, der vil håndtere mad og vand. For eksempel, den kinesiske regering har udstedt love for obligatorisk Salmonella spp. /Shigella spp. screening en gang om året.
Metoden guldstandarden for Salmonella spp. /Shigella spp. opdagelse er bakterier kultur. Gennem bakterier kultur og successive biokemiske identifikation og serologiske karakterisering, kan vi identificere arterne af bakterier, som kan lette sygdom udbrud management og antimikrobiel profilering for at støtte til behandling af patienter 12. det kan også hjælpe med at spore kilden til infektionen i løbet af Salmonella spp. /Shigella spp. udbrud13. Men denne metode er arbejdskrævende (kræver manuel betjening) og tidskrævende (tage flere dage), især for test af et stort antal prøver7. Derudover levedygtige men ikke-bestemmelse (VBNC) Salmonella spp. /Shigella spp.kan findes i nogle afføringsprøver14. I betragtning af disse ulemper, mange laboratorier har forsøgt at udvikle nye teknikker til påvisning af Salmonella spp. /Shigella spp.15,16,17,18 , 19 , 20 , 21 , 22 , 23 , 24 , 25. alle disse metoder bruger nukleare syre forstærkning test (NAAT), blandt hvilke Polymerasekædereaktionen (PCR) er den mest almindelige. En stor begrænsning af disse NAAT baseret metoder er, at døde bakterier, selv bakteriel snavs indeholder ufuldstændige genomisk DNA, kunne vise positive resultater26, som i vid udstrækning kunne påvirke de præcis diagnose af sygdommen. Blanco et al. viste, at Molekylær analyse er meget følsomme, ikke kun til levedygtige Salmonella i kulturer, men også til delvis genomer og døde eller ikke-levedygtige bakterier fra tidligere infektioner eller kontaminering26. Derfor bør der udvikles ny teknologi.
Her, beskrev vi en roman metode at kombinerer NAAT baseret metode og dyrkning. Som vist i figur 1, denne nye metode gælder real-time PCR screening først og derefter positive prøver sendes til bakterier kultur og identifikation.

<table border="0" cellpadding="0" cellspacing="0" style="width:732px;" width="732">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Tris</td>
			<td style="width:112px;">Sigma</td>
			<td style="width:119px;">10708976001</td>
			<td style="width:252px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">EDTA</td>
			<td style="width:112px;">Sigma</td>
			<td style="width:119px;">798681</td>
			<td style="width:252px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">NP40</td>
			<td style="width:112px;">Sigma</td>
			<td style="width:119px;">11332473001</td>
			<td style="width:252px;"></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:249px;">ddH2O</td>
			<td style="width:112px;">Takara</td>
			<td style="width:119px;">9012</td>
			<td style="width:252px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">PrimeSTAR HS (Premix)</td>
			<td style="width:112px;">Takara</td>
			<td style="width:119px;">R040Q</td>
			<td style="width:252px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Nutrient Broth</td>
			<td style="width:112px;">LandBridge</td>
			<td style="width:119px;">CM106</td>
			<td style="width:252px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Nutrient agar</td>
			<td style="width:112px;">LandBridge</td>
			<td style="width:119px;">CM107</td>
			<td style="width:252px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Selenite Cystine medium</td>
			<td style="width:112px;">LandBridge</td>
			<td style="width:119px;">CM225</td>
			<td style="width:252px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">XLD</td>
			<td style="width:112px;">LandBridge</td>
			<td style="width:119px;">CM219</td>
			<td style="width:252px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">MAC&#160;</td>
			<td style="width:112px;">LandBridge</td>
			<td style="width:119px;">CM908</td>
			<td style="width:252px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Salmonella chromogenic agar</td>
			<td style="width:112px;">CHROMagar</td>
			<td style="width:119px;">SA130</td>
			<td style="width:252px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Salmonella diagnostic serum</td>
			<td style="width:112px;">Tianrun</td>
			<td style="width:119px;">SAL60</td>
			<td style="width:252px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Shigella diagnostic serum</td>
			<td style="width:112px;">Tianrun</td>
			<td style="width:119px;">SHI54</td>
			<td style="width:252px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">anal swab (collecting tube plus)</td>
			<td style="width:112px;">Huachenyang</td>
			<td style="width:119px;"></td>
			<td style="width:252px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">slide</td>
			<td style="width:112px;">Mingsheng</td>
			<td style="width:119px;">7102</td>
			<td style="width:252px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">micro-loop</td>
			<td style="width:112px;">Weierkang</td>
			<td style="width:119px;">W511</td>
			<td style="width:252px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">incubator</td>
			<td style="width:112px;">Jinghong</td>
			<td style="width:119px;">DNP-9082</td>
			<td style="width:252px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">autoclave</td>
			<td style="width:112px;">AUL</td>
			<td style="width:119px;">SS-325</td>
			<td style="width:252px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">dry bath</td>
			<td style="width:112px;">Jinghong</td>
			<td style="width:119px;">KB-20</td>
			<td style="width:252px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:249px;">automated microbial identification system</td>
			<td style="width:112px;">bioM&#233;rieux</td>
			<td style="width:119px;">VITEK2</td>
			<td style="width:252px;">other equivalent system could be used</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:249px;">fluorescent real-time PCR machine</td>
			<td style="width:112px;">ThermoFisher</td>
			<td style="width:119px;">ABI7500</td>
			<td style="width:252px;">other equivalent machine could be used</td>
		</tr>
	</tbody>
</table>

a high-throughput platform for the screening of salmonella spp./shigella spp.

Fækal-oral transmission af akut gastroenteritis forekommer fra tid til anden, især når folk, der håndteres fødevarer og vand er smittet med Salmonella spp./Shigella spp. Guld standard metoden til påvisning af Salmonella spp./Shigella spp. er direkte kultur, men dette er arbejdskrævende og tidskrævende. Her beskriver vi en høj overførselshastighed platform for Salmonella spp./Shigella spp. screening, ved hjælp af real-time polymerase kædereaktion (PCR) kombineret med guidet kultur. Der er to større faser: real-time PCR og guidet kultur. For den første fase (real-time PCR) vi forklare hvert trin i metoden: prøve samling, før berigelse, DNA-ekstraktion og real-time PCR. Hvis real-time PCR er positiv, så den anden fase (guidede kultur) udføres: selektiv kultur, biokemiske identifikation og serologiske karakterisering. Vi også illustrere repræsentative resultater, der genereres fra det. Protokollen beskrevet her ville være en værdifuld platform for hurtig, specifikke, følsomme og høj overførselshastighed screening af Salmonella spp. /Shigella spp.

Protokollen blev anvendt til screening af Salmonella spp. /Shigella spp. i anal fæces prøver fra folk, der vil håndtere mad og vand.
I real-time PCR trin, som vist i figur 5A, var der en vellykket forstærkning i HEX kanal, hvilket betød, at den blandede prøve blev testet positiv for Salmonella spp. Så en yderligere real-time PCR blev gennemført på...

Watch this Scientific Journal Video about A High-throughput Platform for the Screening of Salmonella spp./Shigella spp. at JoVE.com

A High-throughput Platform for the Screening of Salmonella spp./Shigella spp.

Salmonella spp. /Shigella spp. er fælles patogener tilskrives diarré. Her, vi beskriver en høj overførselshastighed platform for screening af Salmonella spp. /Shigella spp. ved hjælp af real-time PCR kombineret med guidet kultur.

En høj overførselshastighed Platform for Screening af Salmonella spp. /Shigella spp.

Fecal-oral transmission of acute gastroenteritis occurs from time to time, especially when people who handled food and water are infected by Salmonella ...

Denne metode giver en platform med højt gennemløb til screening af Salmonella og Shigella. Den største fordel ved denne teknik er gentagelse, specifik, følsom og høj gennemløb. Denne teknik kombinerer NAAT hurtig metode og kultur, hvor real-time PCR screening blev anvendt først, og derefter positive blev sendt til bakterier kultur og identifikation.Selv om denne metode fokuserer på Salmonella og Shigella screening, det kan også anvendes på andre patogener såsom Vibrios, Staphylococcus, E.Coli, og et cetera. Visuel demonstration af denne metode er afgørende, da udvælgelsen af den korrekte koloni er vanskelig på grund af baggrundsflora i prøver. Til at begynde, tilføje tre milliliter af næringsstof bouillon til hver prøve.Derefter inkubere prøverne ved 36 grader Celsius i seks timer. Herefter centrifugeres forberedelseskulturen ved 800 g i to minutter. Derefter overføre supernatant til et nyt rør, og centrifuge det igen i fem minutter ved 12, 000 gs.Efter udsmid af supernatant, tilsættes 100 mikroliter DNA-ekstraktionsopløsning til pellet. Så vortex røret i et minut. Dernæst varme prøven ved 1, 000 grader Celsius i det tørre bad.Efter at centrifugering skal supernatanten opsamres i et nyt rør. Først skal du oprette reaktionsblandingen og indstille cykelprogrammet i henhold til tekstprotokollen. Derefter udføre real-time PCR på en fluorescerende real-time PCR maskine.Først tilsættes 100 mikroliter af forberigelseskulturen til fem milliliter selenitglasmedium. Derefter inkubere kulturen ved 36 grader Celsius i 18 til 24 timer. Dernæst indsamle en løkke af kulturen og sprede det på en plade.Derefter inkubere pladen ved 36 grader Celsius i 18 til 24 timer. Efter inkubation skal du vælge mistænkelige kolonier og overføre dem til en plade. Inkuber pladen ved 36 grader Celsius i 18 til 24 timer.Derefter identificere den mistænkelige koloni ved hjælp af et automatiseret mikrobiel identifikationssystem. For at begynde O-antigen karakterisering, tilføje en dråbe af O-antigen polyvalent sera til en ren dias. Overfør en løkke af kolonien til diaset og male det i sera.Hvis bakterierne ligner flydende sand, gentag den tidligere behandling med O-antigen monovalent sera, indtil antigenet er karakteriseret. For at begynde H-antigen karakterisering, tilføje en dråbe H-antigen polyvalent sera til en ren dias. Derefter indsamle en loopful af kolonien og gitter det i sera.Brug H-antigen monovalent sera til at gentage behandlingen, indtil det specifikke H-antigen er karakteriseret. For at adskille kulturen skal du samle en løkke af forberigelseskulturen og sprede den på en plade. Derefter inkubere stedet ved 36 grader Celsius i 18 til 24 timer.Efter inkubation, indsamle en mistænkelig leder koloni på en plade. Inkuber pladen ved 36 grader Celsius i 17 til 24 timer. Derefter udsættes kolonien for biokemisk identifikation ved hjælp af et automatiseret mikrobielt identifikationssystem.Dernæst anvende en dråbe Shigella arter polyvalent sera til en ren dias. Brug derefter en mikro-loop til at indsamle nogle af bakterierne og male det ind i sera. Endelig, hvis sera ligner strømmende sand, bruge Shigella arter monovalent sera at yderligere karakterisere bakterier.Ved hjælp af denne protokol blev afføringsprøver fra patienter screenet for Salmonella- og Shigella-arter. Ved hjælp af PCR i realtid var der en vellykket forstærkning i HEX-kanalen, hvilket indikerer, at prøven var positiv for salmonellaarter. Yderligere pcr i realtid viste, at prøve to var positiv for Salmonella og blev valgt til guidet salmonellaartkultur.I den guidede kultur af Salmonella arter, de lyserøde og lilla kolonier blev belagt separat og underkastet biokemisk identifikation. Resultaterne viste, at kolonierne fra prøve to var Salmonella paratyphi B.Using real-time PCR, der var vellykket forstærkning i FAM-kanalen, hvilket indikerer, at prøven var positiv for Shigella arter. Yderligere PCR i realtid indikerede, at stikprøve 10 var positiv for Shigella og blev valgt til guidet kultur.I shigellaarters guidede kultur blev de lyserøde og farveløse kolonier belagt separat og underkastet biokemisk identifikation. Resultaterne viste, at prøve 10 indeholdt Shigella sonnei fase II bakterier. Mens du forsøger denne procedure, er det vigtigt at huske begrænsningen af metoden på grund af sekvensvariation.Efter denne procedure, andre metoder, ligesom direkte kultur, kan udføres for at undgå falske negative real-time PCR resultater. Denne teknik baner vejen for forskere inden for salmonella- og shigelladetektion, da den nye protokol kan øge den positive hastighed med dobbeltr og mindske arbejdsbyrden og TAT betydeligt. Glem ikke, at arbejde med Salmonella og Shigella kan være ekstremt farligt, og det personlige værnemidler, PPE, bør altid tages, mens du udfører denne procedure.

Research

JoVE Journal

Immunology and Infection

A High-throughput Platform for the Screening of Salmonella spp./Shigella spp.

A Microscopic Phenotypic Assay for the Quantification of Intracellular Mycobacteria Adapted for High-throughput/High-content Screening

En mikroskopisk fænotypisk analyse til kvantificering af intracellulære mycobakterier tilpasset til screening med højt gennemløb/højt indhold

Despite the availability of therapy and vaccine, tuberculosis (TB) remains one of the most deadly and widespread bacterial infections in the world. Since ...

High-throughput Screening for Broad-spectrum Chemical Inhibitors of RNA Viruses

High-throughput Screening for Bredspektret Chemical hæmmere af RNA virus

RNA viruses are responsible for major human diseases such as flu, bronchitis, dengue, Hepatitis C or measles. They also represent an emerging threat ...

High-throughput Assay to Phenotype Salmonella enterica Typhimurium Association, Invasion, and Replication in Macrophages

High-throughput Assay to Phenotype Salmonella enterica Typhimurium Association, Invasion, and Replication in Macrophages

Højkapacitetsforskning Assay til Fænotype<em&gt; Salmonella enterica</em&gt; Typhimurium Association, Invasion, og replikering i makrofager

Salmonella species are zoonotic pathogens and leading causes of food borne illnesses in humans and livestock1. Understanding the mechanisms underlying ...

Establishment of a High-throughput Setup for Screening Small Molecules That Modulate c-di-GMP Signaling in Pseudomonas aeruginosa

Establishment of a High-throughput Setup for Screening Small Molecules That Modulate c-di-GMP Signaling in Pseudomonas aeruginosa

Etablering af en High-throughput Opsætning til screening små molekyler, der modulerer c-di-GMP Signalering i<em&gt; Pseudomonas aeruginosa</em

Bacterial resistance to traditional antibiotics has driven research attempts to identify new drug targets in recently discovered regulatory pathways. ...

Microscopy-based Assays for High-throughput Screening of Host Factors Involved in Brucella Infection of Hela Cells

Microscopy-based Assays for High-throughput Screening of Host Factors Involved in Brucella Infection of Hela Cells

Mikroskopi assays for high-throughput screening af Host faktorer involveret i<em&gt; Brucella</em&gt; Infektion af HeLa-celler

Brucella species are facultative intracellular pathogens that infect animals as their natural hosts. Transmission to humans is most commonly caused by ...

A Fluorescence-based Lymphocyte Assay Suitable for High-throughput Screening of Small Molecules

En Fluorescens-baserede lymfocyt Assay Egnet til high-throughput screening af små molekyler

High-throughput screening (HTS) is currently the mainstay for the identification of chemical entities capable of modulating biochemical reactions or ...

Building Up a High-throughput Screening Platform to Assess the Heterogeneity of HER2 Gene Amplification in Breast Cancers

Opbygge en høj overførselshastighed Screening Platform til at vurdere heterogenitet af HER2 genet forstærkning i brystkræft

Targeted therapies against the human epidermal growth factor receptor 2 (HER2) have radically changed the outcome of patients with HER2-positive breast ...

High-throughput Measurement of Plasma Membrane Resealing Efficiency in Mammalian Cells

Høj overførselshastighed måling af Plasma membran genforsegling effektivitet i pattedyrceller

In their physiological environment, mammalian cells are often subjected to mechanical and biochemical stresses that result in plasma membrane damage. In ...

Chronic Salmonella Infection Induced Intestinal Fibrosis

Kronisk salmonella infektion induceret tarm fibrose

Tissue fibrosis characterized by the pathological accumulation of extracellular matrix such as collagen is the outcome of persistent inflammation and ...