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LIBRIS Formathandbok  (Information om MARC21)
FältnamnIndikatorerMetadata
00005036naa a2200481 4500
001oai:DiVA.org:uu-282792
003SwePub
008160407s2016 | |||||||||||000 ||eng|
0247 a https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-2827922 URI
0247 a https://doi.org/10.1016/j.jcv.2016.01.0152 DOI
040 a (SwePub)uu
041 a engb eng
042 9 SwePub
072 7a ref2 swepub-contenttype
072 7a art2 swepub-publicationtype
1001 a Laiho, Jutta E.u Univ Tampere, Sch Med, Dept Virol, Biokatu 10,FM 3,5th Floor, Tampere 33520, Finland.4 aut
2451 0a Relative sensitivity of immunohistochemistry, multiple reaction monitoring mass spectrometry, in situ hybridization and PCR to detect Coxsackievirus B1 in A549 cells
264 1b Elsevier BV,c 2016
338 a print2 rdacarrier
520 a Background: Enteroviruses (EVs) have been linked to the pathogenesis of several diseases and there is a collective need to develop improved methods for the detection of these viruses in tissue samples. Objectives: This study evaluates the relative sensitivity of immunohistochemistry (IHC), proteomics, in situ hybridization (ISH) and RT-PCR to detect one common EV, Coxsackievirus B1 ( CVB1), in acutely infected human A549 cells in vitro. Study design: A549 cells were infected with CVB1 and diluted with uninfected A549 cells to produce a limited dilution series in which the proportion of infected cells ranged from 10-1 to 10-8. Analyses were carried out by several laboratories using IHC with different anti-EV antibodies, ISH with both ViewRNA and RNAScope systems, liquid chromatography multiple reaction monitoring mass spectrometry (LC/MRM/MS/MS), and two modifications of RT-PCR. Results: RT-PCR was the most sensitive method for EV detection yielding positive signals in the most diluted sample (10-8). LC/MRM/MS/MS detected viral peptides at dilutions as high as 10-7. The sensitivity of IHC depended on the antibody used, and the most sensitive antibody (Dako clone 5D8/1) detected virus proteins at a dilution of 10(-6), while ISH detected the virus at dilutions of 10(-4). Conclusions: All methods were able to detect CVB1 in infected A549 cells. RT-PCR was most sensitive followed by LC/MRM/MS/MS and then IHC. The results from this in vitro survey suggest that all methods are suitable tools for EV detection but that their differential sensitivities need to be considered when interpreting the results from such studies.
653 a Sensitivity
653 a Proteomics
653 a In situ hybridization
653 a Immunohistochemisrty
653 a RT-PCR
7001 a Oikarinen, Maaritu Univ Tampere, Sch Med, Dept Virol, Biokatu 10,FM 3,5th Floor, Tampere 33520, Finland.4 aut
7001 a Richardson, Sarah J.u Univ Exeter, Sch Med, Exeter, Devon, England.4 aut
7001 a Frisk, Gunu Uppsala universitet,Institutionen för immunologi, genetik och patologi4 aut0 (Swepub:uu)gunfrisk
7001 a Nyalwidhe, Juliusu Eastern Virginia Med Sch, Microbiol & Mol Cell Biol, Norfolk, VA 23501 USA.;Eastern Virginia Med Sch, Leroy T Canoles Jr Canc Res Ctr, Norfolk, VA 23501 USA.4 aut
7001 a Burch, Tanya C.u Eastern Virginia Med Sch, Microbiol & Mol Cell Biol, Norfolk, VA 23501 USA.;Eastern Virginia Med Sch, Leroy T Canoles Jr Canc Res Ctr, Norfolk, VA 23501 USA.4 aut
7001 a Morris, Margaret A.u Eastern Virginia Med Sch, Microbiol & Mol Cell Biol, Norfolk, VA 23501 USA.;Eastern Virginia Med Sch, Internal Med, Norfolk, VA 23501 USA.4 aut
7001 a Oikarinen, Samiu Univ Tampere, Sch Med, Dept Virol, Biokatu 10,FM 3,5th Floor, Tampere 33520, Finland.4 aut
7001 a Pugliese, Albertou Univ Miami, Miller Sch Med, Diabet Res Inst, Miami, FL 33136 USA.;Univ Miami, Miller Sch Med, Dept Med, Miami, FL 33136 USA.;Univ Miami, Miller Sch Med, Dept Microbiol, Miami, FL 33136 USA.;Univ Miami, Miller Sch Med, Dept Immunol, Miami, FL 33136 USA.4 aut
7001 a Dotta, Francescou Univ Siena, Diabet Unit, Dept Med Surg & Neurosci, Via Laterina 8, I-53100 Siena, Italy.;Fdn Umberto Di Mario ONLUS Toscana Life Sci, Siena, Italy.4 aut
7001 a Campbell-Thompson, Marthau Univ Florida, Gainesville, FL USA.4 aut
7001 a Nadler, Jerryu Eastern Virginia Med Sch, Internal Med, Norfolk, VA 23501 USA.4 aut
7001 a Morgan, Noel G.u Univ Exeter, Sch Med, Exeter, Devon, England.4 aut
7001 a Hyoty, Heikkiu Univ Tampere, Sch Med, Dept Virol, Biokatu 10,FM 3,5th Floor, Tampere 33520, Finland.;Pirkanmaa Hosp Dist, Fimlab Labs, Tampere, Finland.4 aut
7102 a Univ Tampere, Sch Med, Dept Virol, Biokatu 10,FM 3,5th Floor, Tampere 33520, Finland.b Univ Exeter, Sch Med, Exeter, Devon, England.4 org
7730 t Journal of Clinical Virologyd : Elsevier BVg 77, s. 21-28q 77<21-28x 1386-6532x 1873-5967
8564 u https://europepmc.org/articles/pmc5364806?pdf=render
8564 8u https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-282792
8564 8u https://doi.org/10.1016/j.jcv.2016.01.015

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