Aizhan Tastanova

We confirmed that the melanin-generated color change produced by this biomedical tattoo could be detected with the naked eye and optically quantified. The system was validated in wild-type mice bearing subcutaneously implanted encapsulated engineered cells. All animals inoculated with hypercalcemic breast and colon adenocarcinoma cells developed tattoos, whereas no tattoos were seen in animals inoculated with normocalcemic tumor cells. All tumor-bearing animals remained asymptomatic throughout the 38-day experimental period. Although hypercalcemia is also associated with other pathologies, our findings demonstrate that it is possible to detect hypercalcemia associated with cancer in murine models using this cell-based diagnostic strategy.

We present an optimized dissociation protocol for preparing high-quality skin cell suspensions for in-depth single-cell RNA-sequencing (scRNA-seq) analysis of fresh and cultured human skin. Our protocol enabled the isolation of a consistently high number of highly viable skin cells from small freshly dissociated punch skin biopsies, which we use for scRNA-seq studies. We recapitulated not only the main cell populations of existing single-cell skin atlases, but also identified rare cell populations, such as mast cells. Furthermore, we effectively isolated highly viable single cells from ex vivo cultured skin biopsy fragments and generated a global single-cell map of the explanted human skin. The quality metrics of the generated scRNA-seq datasets were comparable between freshly dissociated and cultured skin. Overall, by enabling efficient cell isolation and comprehensive cell mapping, our skin dissociation-scRNA-seq workflow can greatly facilitate scRNA-seq discoveries across diverse human skin pathologies and ex vivo skin explant experimentations.

Real-time RT-PCR remains a gold standard in the detection of various viral diseases. In the coronavirus 2019 pandemic, multiple RT-PCR–based tests were developed to screen for viral infection. As an emergency response to increasing testing demand, we established a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) PCR diagnostics platform for which we compared different commercial and in-house RT-PCR protocols. Four commercial, one customized, and one in-house RT-PCR protocols were evaluated with 92 SARS-CoV-2–positive and 92 SARS-CoV-2–negative samples. Furthermore, economical and practical characteristics of these protocols were compared. In addition, a highly sensitive digital droplet PCR (ddPCR) method was developed, and application of RT-PCR and ddPCR methods on SARS-CoV-2 environmental samples was examined. Very low limits of detection (1 or 2 viral copies/μL), high sensitivities (93.6% to 97.8%), and high specificities (98.7% to 100%) for the tested RT-PCR protocols were found. Furthermore, the feasibility of downscaling two of the commercial protocols, which could optimize testing capacity, was demonstrated. Tested commercial and customized RT-PCR detection kits show very good and comparable sensitivity and specificity, and the kits could be further optimized for use on SARS-CoV-2 viral samples derived from human and surface swabbed samples. Real-time RT-PCR remains a gold standard in the detection of various viral diseases. In the coronavirus 2019 pandemic, multiple RT-PCR–based tests were developed to screen for viral infection. As an emergency response to increasing testing demand, we established a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) PCR diagnostics platform for which we compared different commercial and in-house RT-PCR protocols. Four commercial, one customized, and one in-house RT-PCR protocols were evaluated with 92 SARS-CoV-2–positive and 92 SARS-CoV-2–negative samples. Furthermore, economical and practical characteristics of these protocols were compared. In addition, a highly sensitive digital droplet PCR (ddPCR) method was developed, and application of RT-PCR and ddPCR methods on SARS-CoV-2 environmental samples was examined. Very low limits of detection (1 or 2 viral copies/μL), high sensitivities (93.6% to 97.8%), and high specificities (98.7% to 100%) for the tested RT-PCR protocols were found. Furthermore, the feasibility of downscaling two of the commercial protocols, which could optimize testing capacity, was demonstrated. Tested commercial and customized RT-PCR detection kits show very good and comparable sensitivity and specificity, and the kits could be further optimized for use on SARS-CoV-2 viral samples derived from human and surface swabbed samples. On March 11, 2020, the World Health Organization (WHO) (Geneva, Switzerland) declared a pandemic because of the quick spread of a respiratory disease caused by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). With cases increasing in multiple countries and high transmissibility of SARS-CoV-2, eradication is rather unrealistic in the short term.1Petersen E. Koopmans M. Go U. Hamer D.H. Petrosillo N. Castelli F. Storgaard M. Al Khalili S. Simonsen L. Comparing SARS-CoV-2 with SARS-CoV and influenza pandemics.Lancet Infect Dis. 2020; 20: e238-e244Abstract Full Text Full Text PDF PubMed Scopus (856) Google Scholar In Switzerland, the second wave of SARS-CoV-2 is predicted to be slower than the first one but with a higher case fatality rate.2Balabdaoui F. Mohr D. Age-stratified discrete compartment model of the COVID-19 epidemic with application to Switzerland.Sci Rep. 2020; 10: 21306Crossref PubMed Scopus (35) Google Scholar The same situation was reported by the WHO for Spanish influenza for which the second and third waves of the infection claimed more lives and the pandemic lasted for almost 2 years and resulted in at least 50 million deaths worldwide [Centers for Disease Control and Prevention (CDC), https://www.cdc.gov/flu/pandemic-resources/1918-commemoration/three-waves.htm, last accessed September 7, 2020]. Another important factor contributing to the rapid spread of the coronavirus disease 2019 (COVID-19) pandemic is an unusually high number of asymptomatic spreaders.3Bi J. Lin Y. Zhong R. Jiang G. Verma V. Shi H. Li J. Tong X. Li Y. Hu D. Liang W. Han G. He J. Prevalence and clinical characterization of cancer patients with asymptomatic SARS-CoV-2 infection history.J Infect. 2020; 81: e22-e24Abstract Full Text Full Text PDF PubMed Scopus (8) Google Scholar,4Oran D.P. Topol E.J. Prevalence of asymptomatic SARS-CoV-2 infection: a narrative review.Ann Intern Med. 2020; 173: 362-367Crossref PubMed Scopus (1373) Google Scholar Therefore, continuous testing and reliable detection of the virus are essential parts of controlling the spread of SARS-CoV-2 (WHO, https://www.who.int/emergencies/diseases/novel-coronavirus-2019/strategies-and-plans, last accessed September 7, 2020). In March 2020, an in-house platform for SARS-CoV-2 diagnostics was initiated as part of an emergency response to an increasing demand for test capacity in a routine microbiology laboratory at University Hospital in Zurich, Switzerland. Currently, the gold standard for the detection and diagnosis of SARS-CoV-2 infection is based on the real-time RT-PCR. The overall goal was to provide in-house SARS-CoV-2 diagnosis to all patients and personnel to ensure the safe and efficient continuation of the health care work within the hospital and the protection of high-risk patients. The aims of this study were to i) evaluate four commercially available, one customized, and one in-house RT-PCR test by comparing the limit of detection (LoD), sensitivity using a panel of SARS-CoV-2 confirmed cases, and specificity using a group of non–COVID-19 respiratory samples; ii) examine the feasibility of down-scaling two commercial protocols to optimize the testing capacity; iii) develop a droplet digital PCR (ddPCR) assay to increase test sensitivity and provide more accurate quantitation of viral RNA; and iv) examine applicability of two validated RT-PCR protocols as well as of a ddPCR protocol on SARS-CoV-2 environmental samples. Patient samples were collected by nasopharyngeal and/or oropharyngeal swabs (CM-FS913, iClean, San Ramon, CA) at the University Hospital Zurich and at ADMed Laboratory in La Chaux-de-Fonds, Switzerland (Copan Diagnostics, Brescia, Italy). The non–COVID-19 samples (other respiratory disease samples) were provided by ADMed Laboratory and were selected after having been tested on the Respiratory Panel FilmArray on Biofire (bioMérieux, Marcy-l'Étoile, France). Household samples were collected by swabbing of the different surfaces in a quarantined household of a SARS-CoV-2–positive patient. All swabs were stored in a viral transport medium (CDC, https://www.cdc.gov/coronavirus/2019-ncov/downloads/Viral-Transport-Medium.pdf, Accessed March 20, 2020) or Eswab (Copan Diagnostics, Murrieta, CA) at 4°C for a maximum of 48 hours or stored at −80°C until further analyses. All household swabbing participants provided informed consent for the study, and both the assay establishment and household studies were approved by the Cantonal Ethics Committee (BASEC-Nr-2020-00660 and BASEC-Nr-2020-00659, respectively). Viral RNA was extracted as previously described5Eichhoff O.M. Bellini E. Lienhard R. Stark W.J. Bechtold P. Grass R.N. Bosshard P.P. Levesque M.P. Comparison of RNA extraction methods for the detection of SARS-CoV-2 by RT-PCR.medRxiv. 2020; https://doi.org/10.1101/2020.08.13.20172494Crossref Scopus (0) Google Scholar using a magnetic bead–based (SpeedBeads, GE Healthcare, Darmstadt, Germany) extraction kit for the KingFisher instrument (MagMax, Thermo Fisher Scientific, Waltham, MA). Four commercially available, one customized (Pasteur Institute, Paris, France), and in-house optimized RT-PCR protocols (Table 1)6Corman V.M. Landt O. Kaiser M. Molenkamp R. Meijer A. Chu D.K. Bleicker T. Brunink S. Schneider J. Schmidt M.L. Mulders D.G. Haagmans B.L. van der Veer B. van den Brink S. Wijsman L. Goderski G. Romette J.L. Ellis J. Zambon M. Peiris M. Goossens H. Reusken C. Koopmans M.P. Drosten C. Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR.Euro Surveill. 2020; 25: 2000045Crossref PubMed Scopus (5072) Google Scholar,7Dong L. Zhou J. Niu C. Wang Q. Pan Y. Sheng S. Wang X. Zhang Y. Yang J. Liu M. Zhao Y. Zhang X. Zhu T. Peng T. Xie J. Gao Y. Wang D. Dai X. Fang X. Highly accurate and sensitive diagnostic detection of SARS-CoV-2 by digital PCR.Talanta. 2021; 224: 121726Crossref PubMed Scopus (97) Google Scholar were compared. Primer probes design, reaction mix, and thermal cycling conditions are given in Table 2, Table 3, Table 4 respectively. All RT-PCR protocols were run according to manufacturer instructions on a QuantStudio 5 DX real-time PCR system (catalog number A36324, Thermo Fisher Scientific), and data were analyzed with the Design and Analysis Software DA version 2.4 (Thermo Fisher Scientific) except for the Euroimmun protocol, which was run on LightCycler 480 II (RocheDiagnostics, Basel, Switzerland). Fast cycling mode was used, and a comparative Ct analysis method was used.Table 1Description of Real-Time RT-PCR Assays Compared in the StudyRT-PCR protocolAbbreviated nameRT-PCR kit/primer and probesMastermix used in this studyPositive controlCDC 2019-Novel Coronavirus Real-Time RT-PCR Diagnostic Panel (for in vitro diagnostics)CDC2019-nCoVEUA-01 Diagnostic Panel Box, catalog number 10006606, IDT, Newark, NJTaMan, Fast Virus 1-step Maste Mix, 4444436, 10 mL, Applied Biosystems/Thermo Fisher Scientific, Waltham, MA2019-nCoV_N_Positive Control, catalog number 10006625, IDTApplied Biosystems TaqMan 2019-nCoV Assay Kit version 1TF-SinglePlexTaqMan 2019-nCoV Assay Kit v1, catalog number A47532, Applied Biosystems/Thermo Fisher ScientificTaMan, Fast Virus 1-step Maste Mix, catalog number 4444436, 10 mL, Applied Biosystems/Thermo Fisher Scientific2019-nCoV Control version 1, catalog number A47533, Applied Biosystems/Thermo Fisher ScientificApplied Biosystems Multiplex TaqMan 2019-nCoV Assay Kit version 2 (research use only)TF-MultiPlexTaqPath COVID-19 Combo Kit, catalog number A47813/A47814, Applied Biosystems/Thermo Fisher ScientificTaqPath1-Step Multiplex Master Mix (No ROX) (4×), catalog number A28523, Applied Biosystems/Thermo Fisher ScientificPositive Control (TaqPath COVID-19 Control Kit), catalog number A47816, Applied Biosystems/Thermo Fisher ScientificEURORealTime SARS-CoV-2 (for research use only)EuroimmunCatalog number MP 2606-0425Provided with the kitProvided with the kitReal-time RT-PCR assays for the detection of SARS-CoV-2,Pasteur Institute, Paris, FrancePasteur Institute Protocol Paris (WHO)https://www.who.int/docs/default-source/coronaviruse/real-time-rt-pcr-assays-for-the-detection-of-sars-cov-2-institut-pasteur-paris.pdf, last accessed November 12, 20206Corman V.M. Landt O. Kaiser M. Molenkamp R. Meijer A. Chu D.K. Bleicker T. Brunink S. Schneider J. Schmidt M.L. Mulders D.G. Haagmans B.L. van der Veer B. van den Brink S. Wijsman L. Goderski G. Romette J.L. Ellis J. Zambon M. Peiris M. Goossens H. Reusken C. Koopmans M.P. Drosten C. Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR.Euro Surveill. 2020; 25: 2000045Crossref PubMed Scopus (5072) Google Scholar; ordered from Microsynth (Balgach, Switzerland)Invitrogen Superscript III Platinum One-Step quantitative RT-PCR system, catalog number 11732-088Available on request from the Pasteur InstituteIn-house customized RT-PCR protocolOncobithttps://www.cdc.gov/coronavirus/2019-ncov/lab/rt-pcr-panel-primer-probes.html, last accessed September 7, 20207Dong L. Zhou J. Niu C. Wang Q. Pan Y. Sheng S. Wang X. Zhang Y. Yang J. Liu M. Zhao Y. Zhang X. Zhu T. Peng T. Xie J. Gao Y. Wang D. Dai X. Fang X. Highly accurate and sensitive diagnostic detection of SARS-CoV-2 by digital PCR.Talanta. 2021; 224: 121726Crossref PubMed Scopus (97) Google Scholar; ordered from MicrosynthTaqPath 1-Step Multiplex Master Mix (no ROX), catalog number A28521, Thermo Fisher ScientificSARS-CoV-2 Positive Run Control, catalog number COV019CE, Bio-Rad, Luxembourg, LuxembourgCDC, Centers for Disease Control and Prevention; nCoV, novel coronavirus; SARS-CoV-2, severe acute respiratory syndrome coronavirus 2; WHO, World Health Organization. Open table in a new tab Table 2Oligonucleotide Sequences of Primers and Probes of Oncobit Real-Time RT-PCR and Digital Droplet PCR ProtocolsPrimer/probe nameSequenceN2 forward primer5′-TTACAAACATTGGCCGCAAA-3′N2 reverse primer5′-GCGCGACATTCCGAAGAA-3′N2 probe (FAM)5′-ACAATTTGCCCCCAGCGCTTCA-3′ORF1ab forward primer5′-CCCTGTGGGTTTTACACTTAA-3′ORF1ab reverse primer5′-ACGATTGTGCATCAGCTGA-3′ORF1ab probe (Cy5)5′-CCGTCTGCGGTATGTGGAAAGGTTATGG-3′RNaseP forward primer5′-AGATTTGGACCTGCGAGCG-3′RNaseP reverse primer5′-GAGCGGCTGTCTCCACAAGT-3′RNaseP probe (HEX)5′-TTCTGACCTGAAGGCTCTGCGCG-3′ Open table in a new tab Table 3Reaction Mix for Oncobit Real-Time RT-PCR ProtocolReagentVolume per reaction, μLTaqPath 1-Step Multiplex Master Mix (no ROX) (catalog number A28521, Thermo Fisher Scientific, Waltham, MA), 4×5N2 probe (FAM) (100 μmol/L)0.05ORF1ab probe (Cy5) (100 μmol/L)0.05RNaseP probe (HEX) (100 μmol/L)0.05N2 forward primer (100 μmol/L)0.06N2 reverse primer (100 μmol/L)0.06ORF1ab forward primer (100 μmol/L)0.06ORF1ab reverse primer (100 μmol/L)0.06RNaseP forward primer (100 μmol/L)0.03RNaseP reverse primer (100 μmol/L)0.03Nuclease-free water4.55Total20.0 Open table in a new tab Table 4Thermal Cycling Conditions for Oncobit Real-Time RT-PCR ProtocolStageStepTemperature, °CTimeHoldUracil-DNA glycosylase incubation252 minutesHoldReverse transcription5310 minutesHoldActivation952 minutesCycling (40 cycles)Denaturation953 secondsAnneal/extension6030 seconds Open table in a new tab CDC, Centers for Disease Control and Prevention; nCoV, novel coronavirus; SARS-CoV-2, severe acute respiratory syndrome coronavirus 2; WHO, World Health Organization. For the CDC protocol, an RT-PCR result was defined as inconclusive if only the N1 gene (±N3 gene) was positive or if only the N2 gene (±N3 gene) was positive. For the TF-MultiPlex (Thermo Fisher Scientific), TF-SinglePlex (Thermo Fisher Scientific), and Oncobit protocols, an RT-PCR result was considered inconclusive if only one of the two or three of the viral genes was positive. Inconclusive results were not repeated. The Euroimmun protocol (Luebeck, Germany) does not have the inconclusive category. The ddPCR protocol for SARS-CoV-2 detection targets two viral genomic regions of the SARS-CoV-2 gene (ORF1ab and N2) and uses the human RNase P gene as an in-process control. The following probes for the three genes were used: ORF1ab (FAM and HEX), N2 (FAM), and RNase P (HEX) (Table 2). Briefly, 20 μL of reaction mix (containing 1-Step RT-ddPCR Advanced Kit for Probes Mastermix; Bio-Rad, Luxembourg, Luxembourg) was combined with 10 μL of RNA sample for a final reaction volume of 30 μL. The final concentrations were 90 nmol/L for primers (ORF1ab, N2, RNaseP), 19.5 nmol/L for RdRP probes, 30 nmol/L for the N2 probe, and 40 nmol/L for the RNase P probe. The SARS-CoV-2 Positive Run Control (catalog number COV019CE, Bio-Rad) was used as positive control. ddPCR was run according to the program listed in Table 5 using QX200 Droplet Digital PCR System (Bio-Rad). The swabbing household samples from a laptop, newspaper, or door handle as well as the nontemplate control were tested in two independent Cycling Conditions for Oncobit Digital Droplet PCR minutesCycling Open table in a new tab The of four SARS-CoV-2 detection protocols (CDC, and was using a of an quantitative test sample last accessed November 12, 2020). was used to the of for the Ct and viral Ct of 40 was as the of viral by RT-PCR. for Oncobit ddPCR protocol was using a of the SARS-CoV-2 Positive Run Control (catalog number COV019CE, For sensitivity and specificity of the results of RT-PCR from the ADMed Laboratory were used as the gold standard The sensitivity was defined with the specificity was defined as and the result of tested assays the was the result from the tested assays not the was Inconclusive results were from sensitivity and specificity The viral assay was as previously C. M. H. and tests for and Google F. Wang A. Liu M. Wang Q. J. S. Y. Zhang Y. J. L. Jiang S. H. Y. J. to SARS-CoV-2 in a COVID-19 and 2020; Scopus (0) Google H. H. S. Wang Y. Hu B. X. Y. Yang D. Y. Zhang X. Chu Zhou J. and of SARS-CoV-2 and SARS-CoV with for clinical and laboratory studies of an 2020; Full Text Full Text PDF PubMed Google Scholar with Briefly, 5 (catalog number were on in μL of high medium with and (catalog number France). hours of the medium was and μL of a virus test or the positive SARS-CoV-2 control by University of Switzerland) was in to the The were for 48 hours at The were with for at with and with (catalog number Pan Darmstadt, Germany) for at The was the were with and the at for viral The RT-PCR protocols compared in this study use the same of viral RNA from the nasopharyngeal and/or oropharyngeal swabs or and a 1-step reaction by real-time of two or three SARS-CoV-2 genes and of all tested RT-PCR protocols is given in Table All protocols have nontemplate and positive In the is as the control as both RNA and reaction control. All protocols except for Euroimmun the of control is not use a reaction control to ensure RNA and RT-PCR reaction was not The protocol is or Euroimmun protocol with design, with two probes to the same The viral RNA is 5 to 10 μL. of Table the protocol developed by Pasteur Institute was not used further in this comparative of RT-PCR Institute Protocol genes and gene per volume per reaction volume per reaction Centers for disease control and SARS-CoV-2, severe acute respiratory syndrome coronavirus 2; WHO, World Health Organization. Open table in a new tab CDC, Centers for disease control and SARS-CoV-2, severe acute respiratory syndrome coronavirus 2; WHO, World Health Organization. With a Ct of the RT-PCR SARS-CoV-2 detection protocols (CDC, and as well as the Oncobit ddPCR protocol an and 2 viral 2, and high sensitivity of the tested protocol 2, and For the sensitivity and specificity of the SARS-CoV-2 detection protocols (CDC, and a of 92 SARS-CoV-2–positive samples and 92 SARS-CoV-2–negative samples was used were provided by ADMed to SARS-CoV-2–positive results sensitivity of all tested protocols, with a for TF-SinglePlex and to sensitivity for the protocols In the specificity samples a confirmed diagnosis of respiratory Table and samples tested for all listed respiratory All protocols, except samples tested positive for four of Table The specificity was for all protocols except for which a specificity of Inconclusive results were in to of these with TF-MultiPlex and Oncobit the Comparing RT-PCR results or of all the overall all the protocols was optimize testing capacity, reaction in commercial protocols were and were customized to have an in-house developed protocol a previously confirmed SARS-CoV-2–positive of the CDC and TF-MultiPlex protocols were compared with reaction volume and reaction by RNA sample was the The Oncobit protocol was the (Table the RT-PCR reaction (Table and the reliable to Switzerland). The specificity and sensitivity of the Oncobit protocol were comparable with commercial SARS-CoV-2 RT-PCR detection kits 3, and CDC protocol two inconclusive a standard TF-MultiPlex protocol one and a TF-MultiPlex protocol one as well as one inconclusive result Furthermore, of CDC and TF-MultiPlex protocols a sensitivity of with a Ct of 40 compared and established the RT-PCR protocols for SARS-CoV-2 the of application of the RT-PCR and ddPCR protocol for SARS-CoV-2 detection on environmental samples was examined. of different surfaces from a SARS-CoV-2 quarantined household were collected and analyzed by two validated RT-PCR protocols. In addition, an in-house ddPCR protocol was developed to and On the of household surface swabbing 2020) of the SARS-CoV-2–positive only 2 was swabbed and tested positive but reported The as well as the swabbed surface samples were collected on the same and tested with three different SARS-CoV-2 detection protocols (CDC, and The tested positive on three different protocols. The and two more swabbed surface door handle and samples positive and inconclusive Table for of the samples was Real-time RT-PCR remains the sensitive method for detection of a of specificity, and practical of four commercial SARS-CoV-2 detection kits as well as one optimized in-house RT-PCR SARS-CoV-2 study comparing RT-PCR with rapid SARS-CoV-2 detection method sensitivity of the rapid method was only B. Zhang J. J. Han C. Y. Wang S. G. Zhou H. Y. of a rapid test in the diagnosis of SARS-CoV-2 Infect. 2021; Full Text Full Text PDF PubMed Scopus Google Scholar; RT-PCR remains a more sensitive detection method for of the reported studies on real-time RT-PCR SARS-CoV-2 detection the only on a number of samples and tested only commercial detection M. B. V. J. Koopmans M. Molenkamp R. Comparison of commercial reverse PCR assays for the detection of 2020; PubMed Scopus Google van der Veer B. van den Brink S. Wijsman L. J. van den A. Molenkamp R. Reusken C. Meijer A. Comparison of commercial RT-PCR diagnostic kits for 2020; PubMed Scopus Google Scholar and studies the only to sensitivity W. R. E. Comparison of four in vitro diagnostic assays for the detection of SARS-CoV-2 in nasopharyngeal 2020; PubMed Scopus Google Scholar In this study, a low and high sensitivity for four commercial SARS-CoV-2 RT-PCR detection protocols were by using standard quantitative test samples and a of 92 SARS-CoV-2–positive respectively. Furthermore, specificity of protocols was tested and confirmed with 92 samples confirmed SARS-CoV-2–negative result or were collected in from patients with respiratory Table In addition, downscaling of two commercial protocols were for the diagnostic routine and could be an to downscaling is important in a high demand for SARS-CoV-2 testing as at the of the pandemic in As an to optimize and increase testing capacity, an in-house protocol was developed in with the diagnostics Oncobit by previously primer for The customized Oncobit protocol was the least and protocol compared with commercial RT-PCR protocols tested in this the application of RT-PCR–based detection protocols, a testing of swabbed surfaces from a quarantined household was RT-PCR protocols the viral on the and this result was confirmed by a more sensitive ddPCR more surfaces inconclusive results and a door with viral by the Table on the same tested assay for all samples the of the RT-PCR–based protocols on samples could be of use for environmental the comparative study, we commercial and customized RT-PCR–based detection protocols are highly at viral in nasopharyngeal and/or oropharyngeal and because of high RT-PCR–based detection protocols be to the testing of environmental samples. for with for the study, and and the of University Hospital Zurich for with the and Institute of and University of Switzerland for positive SARS-CoV-2 control for The of the study was by and were by and The were and were analyzed by and The study was by and The was by and All the final with Ct and as well as of swabbed and surface samples. The Ct and of Ct of the N2, and viral genes are for Disease Control and Prevention Patient was a 2, 3, and patients The Ct at the of diagnosis for are because the was tested in a different On 7, 2020, the of which lasted for On 7, 2020, the tested positive for severe acute respiratory syndrome coronavirus 2 with after 10 As the we increasing Ct in both patients. On 12, the two and tested positive for SARS-CoV-2 but asymptomatic at all in the household were swabbed on the with patients asymptomatic for at least 2 assay using for of the samples tested positive by real-time RT-PCR newspaper, and with Table with Table with Table

Immune checkpoint inhibitors are standard-of-care for the treatment of advanced melanoma, but their use is limited by immune-related adverse events. Proteomic analyses and multiplex cytokine and chemokine assays from serum at baseline and at the adverse event onset indicated aberrant T cell activity with differential expression of type I and III immune signatures. This was in line with the finding of an increase in the proportion of CD4+ T cells with IL-17A expression at the adverse event onset in the peripheral blood using flow cytometry. Multiplex immunohistochemistry and spatial transcriptomics on immunotherapy-induced skin rash and colitis showed an increase in the proportion of CD4+ T cells with IL-17A expression. Anti-IL-17A was administered in two patients with mild myocarditis, colitis and skin rash with resolution of the adverse events. This study highlights the potential role of type III CD4+ T cells in adverse event development and provides proof-of-principle evidence for a clinical trial using anti-IL-17A for treating adverse events. Dimitriou et al. perform multiomic profiling of patients with melanoma experiencing immunotherapy-associated toxicity and identify a targetable role for type III-associated immune responses with an increase in CD4+ T cells expressing IL-17A.