Cães Detectores de COVID-19-Identificação de Amostras por Olfato-Artigo Científico

RESEARCH ARTICLE Open Access Scent dog identification of samples from COVID19 patients a pilot study Paula Jendrny1 Claudia Schulz2 Friederike Twele1 Sebastian Meller1 Maren von KöckritzBlickwede23 Albertus Dominicus Marcellinus Erasmus Osterhaus2 Janek Ebbers4 Veronika Pilchová2 Isabell Pink5 Tobias Welte5 Michael Peter Manns6 Anahita Fathi789 Christiane Ernst10 Marylyn Martina Addo789 Esther Schalke11 and Holger Andreas Volk1 Abstract Background As the COVID19 pandemic continues to spread early ideally realtime identification of SARSCoV2 infected individuals is pivotal in interrupting infection chains Volatile organic compounds produced during respiratory infections can cause specific scent imprints which can be detected by trained dogs with a high rate of precision Methods Eight detection dogs were trained for 1 week to detect saliva or tracheobronchial secretions of SARSCoV2 infected patients in a randomised doubleblinded and controlled study Results The dogs were able to discriminate between samples of infected positive and noninfected negative individuals with average diagnostic sensitivity of 8263 95 confidence interval CI 82028324 and specificity of 9635 95 CI 96319639 During the presentation of 1012 randomised samples the dogs achieved an overall average detection rate of 94 34 with 157 correct indications of positive 792 correct rejections of negative 33 incorrect indications of negative or incorrect rejections of 30 positive sample presentations Conclusions These preliminary findings indicate that trained detection dogs can identify respiratory secretion samples from hospitalised and clinically diseased SARSCoV2 infected individuals by discriminating between samples from SARSCoV2 infected patients and negative controls This data may form the basis for the reliable screening method of SARSCoV2 infected people Keywords COVID19 SARSCoV2 Volatile organic compounds Scent detection dogs Olfactory detection Saliva Background The ongoing COVID19 pandemic highlights the import ance of fast and reliable testing for accurate identification of symptomatic and asymptomatic carriers to reduce spread of infection effectively 1 Current testing regimens usually require nasopharyngeal swabs applied by a trained person and a reverse transcription polymerase chain reaction test RTPCR for pathogen identification Obtaining RTPCR results is time consuming and can be costprohibitive espe cially for developing countries and is therefore currently often used in a targeted fashion testing predominantly pa tients with COVID19 specific symptoms 1 There is therefore a need for an additional faster reliable non invasive and versatile screening tool especially to identify asymptomatic and presymptomatic individuals Several studies have proven the canines extraordinary olfactory acuity to detect persons with infectious and noninfectious diseases like different types of cancer 2 malaria 3 bacterial and viral infections 46 with usually high rates of sensitivity and specificity 7 A pathogenspecific odour that can be detected by dogs The Authors 2020 Open Access This article is licensed under a Creative Commons Attribution 40 International License which permits use sharing adaptation distribution and reproduction in any medium or format as long as you give appropriate credit to the original authors and the source provide a link to the Creative Commons licence and indicate if changes were made The images or other third party material in this article are included in the articles Creative Commons licence unless indicated otherwise in a credit line to the material If material is not included in the articles Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use you will need to obtain permission directly from the copyright holder To view a copy of this licence visit httpcreativecommonsorglicensesby40 The Creative Commons Public Domain Dedication waiver httpcreativecommonsorgpublicdomainzero10 applies to the data made available in this article unless otherwise stated in a credit line to the data Correspondence holgervolktihohannoverde Claudia Schulz and Friederike Twele contributed equally to this work 1Department of Small Animal Medicine and Surgery University of Veterinary Medicine Hannover Hannover Germany Full list of author information is available at the end of the article Jendrny et al BMC Infectious Diseases 2020 20536 httpsdoiorg101186s12879020052813 may be composed of specific patterns of volatile organic compounds VOCs Compared to bacteria viruses have no own metabolism and therefore VOCs are released by infected body cells as a result of metabolic host pro cesses 8 Different technical approaches have used the detection of VOCs to discriminate infectious diseases successfully but none is being used routinely in clinical practice 9 As dogs can be trained quickly the aim of the present study was to test the concept of using dogs reliably and in realtime to discriminate between sam ples of SARSCoV2 infected patients and noninfected controls This method could be employed in public areas such as airports sport events borders or other mass gatherings as an alternative or addition to laboratory testing thus helping to prevent further spreading of the virus or further outbreaks Methods Sample acquisition Saliva samples and tracheobronchial secretion samples were collected from hospitalised COVID19 patients that showed clinical symptoms and were diagnosed as SARS CoV2 positive using nasopharyngeal swab samples Negative control samples were obtained from SARS CoV2 RTPCR negative people with no previous history of COVID19 nor had the individuals any history of a recent cold or infection None of the samples were screened for different human coronaviruses like beta coronavirus HCoVOC43 or alpha coronavirus HCoV 229E After the sample acquisition the anonymised sam ples were transported to the University of Veterinary Medicine Hannover Sample preparation All collected samples were confirmed as positive or negative using the RTPCR SARSCoV2IP4 assay from Institut Pasteur recommended by the World Health Organization 10 11 including an internal control sys tem and protocol as described 12 13 Samples from COVID19 patients irrespective of the final RTPCR re sult were further subjected to virus quantification end point dilution assay and virus isolation analysis using Vero E6 cells under biosafety level 3 conditions The cell layers were assessed for cytopathic effects and final re sults were obtained 7 days after cell infection Since dogs are susceptible to SARSCoV2 14 all samples from COVID19 patients were inactivated using beta propio lactone BPL in order to protect the dogs and their han dlers from infection during training Briefly samples and reagents were kept at 4 C 20 μlml NaHCO3 75 was added and samples were incubated for 10 min at 4 C After addition of 10 μlml of 10 BPL samples were in cubated at 4 C for 70 to 72 h Hydrolysis of BPL was conducted at 37 C for 1 to 2 h Samples that showed a cytopathic effect before BPL inactivation using virus iso lation or end point dilution assay were tested again after BPL inactivation and were confirmed to be inactivated Only BPL inactivated samples from COVID19 patients were used for the dog training Furthermore detection dogs were provided both negative control samples with and without previous BPL treatment to exclude hydro lysed BPL as a potential distracting reagent For the dog training a volume of 100 μl per sample was pipetted onto a cotton pad which was placed into a 4 ml glass tube Dog training and study design The presentation of the samples to the dogs was con ducted via a device called Detection Dog Training System DDTS Kynoscience UG Germany which can present samples in a randomised automated manner without trainer interference For a short video sequence see Add itional file 1 DDTS was utilised for training and testing The device is composed of seven scent holes Behind each hole two tubes are leading to two metal containers In the study the first container enclosed the target sample and the second one carried the control sample Only one con tainer is presented in each sniffing hole at any given time as the pairs of containers are situated on movable slides inside the device The metal containers were covered with grids which allowed the odour to escape and reach the sniffing hole Each tube extension was identical and L shaped which prevented dogs from physical contact with the samples and excluded any visual cues that may have enabled further detection capabilities For each trial run only one hole presented a SARSCoV2 positive sample at a time while the other six holes presented negative sam ples After the indication of the hole with the positive sample the dog was automatically rewarded by the device with food or ball The indication time was changed during successful training from 1 s to 2 s While the reward was eaten the devices software randomly and automatically assigned new positions to the slides for the following ses sion with again only one hole presenting the positive odour sample The dog its handler and a person observing the study were blinded during the doubleblinded study All personnel stood behind the dog during the test runs to avoid distraction The device recorded automatically the number and time length of each nose dip into the scent holes and the location of the positive and negative samples This was verified by manual timestamped video analysis Analysis of sensitivity and specificity The diagnostic sensitivity Se true positive TP TP false negative FN diagnostic specificity Sp true negative TN TN false positive FP positive pre dictive values PPV TPTP FP and negative Jendrny et al BMC Infectious Diseases 2020 20536 Page 2 of 7 predictive values NPV TNTN FN were calculated according to Trevethan 15 Results After a 2 weeks habituation process to the DDTS the eight dogs needed 5 days of training in total until the de tection rate was above chance An additional spreadsheet provides background information of the dogs used in the study see Additional file 2 The controlled double blinded detection study was then conducted after 7 days of training and in total 10388 sample presentations Table 1 On each training day unknown and known positive samples and negative control samples were presented to the canines The response to the new sample was used in order to evaluate if the generalisation process has been achieved While the dogs had only achieved an average detection rate of 50 on the second day of train ing the values increased to 70 on day five and even 81 on day seven indicating a successful generalisation process After completion of the training process the de tection accuracy of the eight trained dogs was evaluated in a randomised doubleblinded and controlled study Table 2 Samples from seven infected and seven healthy individuals were used in this study Two of the positive samples were tracheobronchial secretion the other samples consisted of saliva Within randomised and automated 1012 sample pre sentations dogs achieved an overall average detection rate of 94 34 with 157 correct indications of posi tive 792 correct rejections of negative 33 false positive and 30 false negative indications The canines discrimi nated between infected and noninfected individuals with an overall diagnostic sensitivity of 8263 95 confidence interval CI 82028324 and specificity of 9635 95 CI 96319639 Sensitivity ranged from 679 to 952 and specificity from 924 to 989 Fig 1 There was no notable difference in detection ability be tween saliva and tracheal secretion average hit rates 851 and 877 respectively Discussion Timely and accurate detection of SARSCoV2 infected individuals is of uttermost importance for a society to control the pandemic Our data indicate that detection dogs can be trained in just about a week to discriminate between samples of people infected and noninfected by SARSCoV2 The average detection rate was 94 Ana lysis for accuracy and precision revealed a diagnostic sensitivity of 8263 95 CI 82028324 and a high diagnostic specificity of 9635 95 CI 96319639 for all dogs All dogs had a high diagnostic specificity with a small range in variation which could be import ant for population screening to avoid false positive re sults However there was quite a range in variation of sensitivity for the individual dog and inbetween dogs This can in part be explained with the dogs variable training background see Additional file 2 signalment personality traits and short training period of 7 days To avoid a bias concerning hospital specific smells positive samples were obtained from two different hospitals to include a variation in a covariate factor and this appears to have not influenced the current results Understand ing better why there is this range in sensitivity and how to potentially improve it would be important prior to considering the use of detection dogs in the field In Table 1 Number of presented samples per dog during training Day Sample status Dog 1 Dog 2 Dog 3 Dog 4 Dog 5 Dog 6 Dog 7 Dog 8 Total 1 positive 15 20 15 15 15 15 15 10 120 negative 90 120 90 90 90 90 90 60 720 2 positive 15 15 15 15 15 15 10 15 115 negative 90 90 90 90 90 90 60 90 690 3 positive 35 35 35 35 30 35 38 35 278 negative 210 210 210 210 180 210 228 210 1668 4 positive 20 15 20 20 20 40 60 20 215 negative 120 90 120 120 120 240 360 120 1290 5 positive 40 40 30 30 35 30 53 60 318 negative 240 240 180 180 210 180 318 360 1908 6 positive 20 7 20 14 10 10 30 15 126 negative 120 42 120 84 60 60 180 90 756 7 positive 50 30 50 47 30 30 35 40 312 negative 300 180 300 282 180 180 210 240 1872 Total samples 1365 1134 1295 1232 1085 1225 1687 1365 10388 Jendrny et al BMC Infectious Diseases 2020 20536 Page 3 of 7 Table 2 Diagnostic performance of the eight scent detection dogs Detect ion by dog SARSCoV2 infection status Total number Diagnostic specificity Sp Diagnositic sensitivity Se Standard Error SE Sp Standard Error SE Se Confidence Interval 95CI Sp Confidence Interval 95CI Se Negative predictive value NPV Positive predictive value PPV Standard Error SE NPV Standard Error SE PPV Confidence Interval 95CI NPV Confidence Interval 95CI PPV negative positive Dog 1 No 89 3 113 099 087 001 007 0002 0029 097 095 002 004 0004 0018 Yes 1 20 Dog 2 No 91 1 115 097 095 002 005 0004 0020 099 087 048 007 0097 0031 Yes 3 20 Dog 3 No 81 2 104 099 091 001 006 0003 0026 098 095 062 005 0134 0019 Yes 1 20 Dog 4 No 95 4 120 097 082 002 008 0003 0034 096 086 069 007 0136 0031 Yes 3 18 Dog 5 No 110 2 135 097 091 002 006 0003 0026 098 087 053 007 0099 0030 Yes 3 20 Dog 6 No 137 8 170 096 071 002 009 0003 0032 094 080 078 008 0129 0028 Yes 5 20 Dog 7 No 92 5 122 095 080 002 008 0004 0031 095 080 078 008 0156 0031 Yes 5 20 Dog 8 No 97 8 133 092 070 003 009 0005 0033 092 068 089 009 0170 0034 Yes 9 19 Total No 792 33 1012 096 083 001 003 00004 0004 096 084 118 003 0081 0004 Yes 30 157 Jendrny et al BMC Infectious Diseases 2020 20536 Page 4 of 7 comparison the current gold standard diagnostic RT PCR test of a nasopharyngeal swab can in trained hands have a false detection rate of 25 and a false positive rate of 2369 16 A new not yet published study in dicated a clear nearly 100 VOC specific pattern of SARSCOV2 infected individuals compared to negative controls and individuals infected by the influenza virus using multicapillary column coupled ion mobility spec trometry of breath 17 This provides further indications that unique VOC imprints exist and can be used for the development of diagnostic procedures The current study results are promising although they should be regarded as preliminary and suitability for this detection method in the field can only be acquired after further research has been conducted Our work provides the very first steps of the development of a new SARS CoV2 screening method Our inclusion criteria for the samples collected were rather nonspecific and not stratified by severity of symptoms disease status or virus load Future studies are needed to address this including a higher number of different samples to evaluate the analytical sensitivity eg dilution of samplesdetection level different disease phenotypes and stages and ana lytical specificity differentiation to other lung diseases or pathogens such as cancer or infection with other sea sonal respiratory virus infections eg influenza respira tory syncitial virus adenovirus other than SARSCoV2 coronaviruses rhinovirus In the current study negative control samples were acquired from healthy individuals without clinical signs of respiratory disease The individ uals were only tested for SARSCoV2 virus and there fore one cannot exclude that a former infection especially with another human coronavirus like HCoV OC43 resulted in false positive indications of the dogs and that cross detection occurred On the other hand samples included in the current study were from se verely affected hospitalised COVID19 patients but one of the main challenges in controlling the current pan demic is to identify presymptomatic COVID19 patients and asymptomatic carriers which may constitute most COVID19 cases 18 The sensitivity of detection by dogs may also vary across the course of the disease Fu ture research should therefore focus on the ability of dogs to identify the different COVID19 disease pheno types and phases of disease expression such as asymp tomatic presymptomatic mild and severe clinical cases as well as to test samples of the same individuals at dif ferent timepoints across the course of the disease One of the most important requirements regarding hand ling of infectious samples is infection prevention and con trol Initially it was assumed that dogs cannot get infected by SARSCoV2 but recent single cases have been reported showing that dogs can get infected by SARSCoV2 and could potentially play a role in viral spread 14 19 There is evidence of humantoanimal transmission with a subse quent infection of dogs It is still unclear whether dogs can function as spreaders of the virus by infecting other animals or humans 14 20 Nevertheless this needs to be consid ered when using dogs for detection of infected material or people It is also unclear how an infection in the dog will alter its sense of smell In the current study we chose to use an inactivation procedure which should not affect VOCs However this is not practical for testing in the field and we are currently developing new strategies for a secure presen tation of noninactivated samples This would eliminate po tential risks of virus transmission by detection dogs when used in a nonlaboratory setting Conclusions Detection dogs were able to discriminate respiratory secre tions of infected SARSCoV2 individuals from those of healthy controls with high rates of sensitivity and specificity The current pilot study had major limitations which needs to be elucidated in future studies SARSCoV2 detection dogs may then provide an effective and reliable infection detection technology in various settings like public facilities and func tion as an alternative or addition to regular RTPCR screen ing In countries with limited access to diagnostic tests detection dogs could then have the potential to be used for mass detection of infected people Further work is necessary to better understand the potential and limitation of using scent dogs for the detection of viral respiratory diseases Supplementary information Supplementary information accompanies this paper at httpsdoiorg10 1186s12879020052813 Additional file 1 Additional video Detection dog working with DDTS The video Additional file 1 shows the Labrador Retriever Seven Fig 1 Diagnostic specificity and sensitivity by dog and for all dogs together Whiskers show 95 confidence intervals Jendrny et al BMC Infectious Diseases 2020 20536 Page 5 of 7 during a detection session The Detection Dog Training System DDTS can be seen at the bottom of the video The scent hole with a sample of an SARSCoV2 infected individual is marked in green on the video please note the green mark was not seen by the dog and was only used in the video as a visualisation tool for the viewer to demonstrate the dogs search and detection behaviour At each detection trial run only one hole is presenting the target scent with the other six holes present ing saliva samples from SARSCoV2 negative tested individuals When the dog detects the target scent the nose will be left within the hole for 2 s to indicate the detection This will be recorded by the device A beeping sound announces the food or ball reward which is automatically ejected by the device distracting the dog for a short time period In the meantime the device rearranges the sample presentation in an auto matic and random fashion presenting one other scent hole with a sam ple of a SARSCoV2 positive tested individual and six control scent holes with negative control samples In the upper left corner of the video one can see how the figures change depending on the detection behaviour of the dog true positive correct indication n 3 true negative correct rejection n 8 false positive incorrect indication n 0 and false nega tive incorrect rejection n 1 Additional file 2 Additional Table Characteristics of the dogs The additional table Additional file 2 shows the signalment and background of the eight dogs that participated in the study Abbreviations RTPCR reverse transcription polymerase chain reaction test BPL beta propiolactone VOCs volatile organic compounds DDTS Detection Dog Training System CI confidence interval TP true positive FN false negative Sp specificity TN true negative FP false positive PPV positive predictive values NPV negative predictive values Acknowledgements We would like to thank the members of the IDUKECOVID19 Study Group Marylyn M Addo Etienne Bartels Thomas T Brehm Christine Dahlke Anahita Fathi Monika Friedrich Svenja Hardtke Till Koch Ansgar W Lohse My L Ly Stefan Schmiedel L Marie Weskamm Julian Schulze zur Wiesch at the Uni versity MedicalCenter HamburgEppendorf by helping us with recruitment of patients and sample collection We further would like to thank Stefan Hampel Sina Knisel and Miguel Acosta for support at the Bundeswehr School of Dog German Armed Forces Ulmen during dog training and Lean der Buchner from the Central Institute of Medical Services German Armed Forces in Koblenz for the support in getting samples A special thanks goes to Hans Ebbers Kynosciences for providing the DDTS free of charge and for the support in dog training We would like to thank our doctoral student Saskia Irene Peek for her support during sample collection Special thanks go to our doggy noses Coyote Elli Lotta Donnie Hoss Luigi Jo and Seven Heartfelt thanks go to all the people providing us with samples especially to the SARSCoV2 infected persons and their relatives with the sincere intention to contribute to the containment of COVID19 and to scientific pro gress We wish you lots of strength and full recovery during the current pandemic Authors contributions PJ participated in the planning of the study carried out the main practical work data analyses and drafted the manuscript CS participated in the planning of the study and carried out the laboratory work including RTPCR and virus inactivation as did VP FT SM and HAV designed and coordinated the study drafted the manuscript conducted and coordinated FT the sam ple acquisition and were responsible for data analyses MvKB and ADMEO participated in the planning of the laboratory part of the study and were in charge for the legal permission for sample processing JE programmed the DDTS software IP TW MPM AF and MMA were in charge for the ethical ap proval patient recruitment and sample collection IP AF at Hannover Med ical School IP TW MPM and University MedicalCenter HamburgEppendorf AF MMA ES participated in the planning of the study was responsible for the dog training and helped with data analyses All authors have read and approved the final manuscript Funding Not applicable Availability of data and materials The datasets used andor analysed during the current study are available at Jendrny Paula Twele Friederike Schulz Claudia Meller Sebastian von KöckritzBlickwede Maren Volk Holger Andreas 2020 SARSCoV2 detec tion dogs a pilot study Data set Zenodo httpsdoiorg105281zenodo 3950074 Ethics approval and consent to participate The study was conducted according to the ethical requirements established by the Declaration of Helsinki The local Ethics Committee of Hannover Medical School MHH and Hamburg Medical Association at the University MedicalCenter HamburgEppendorf UKE approved the study ethic consent number 9042BOK2020 and PV7298 respectively Written consent from all people were collected before sample collection Consent for publication Not applicable Competing interests The authors declare that they have no competing interests Author details 1Department of Small Animal Medicine and Surgery University of Veterinary Medicine Hannover Hannover Germany 2Research Center for Emerging Infections and Zoonoses University of Veterinary Medicine Hannover Hannover Germany 3Department of Physiological Chemistry University of Veterinary Medicine Hannover Hannover Germany 4Hörstel Germany 5Department of Respiratory Medicine Hannover Medical School Hannover Germany 6Hannover Medical School Hannover Germany 7Department of Medicine Division of Infectious Diseases University MedicalCenter HamburgEppendorf Hamburg Germany 8Department for Clinical Immunology of Infectious Diseases Bernhard Nocht Institute for Tropical Medicine Hamburg Germany 9German Center for Infection Research HamburgLübeck BorstelRiems Germany 10Central Institute of Medical Service German Armed Forces Koblenz Germany 11Bundeswehr School of Dog handling German Armed Forces Ulmen Germany Received 16 July 2020 Accepted 21 July 2020 References 1 Wiersinga WJ Rhodes A Cheng AC Peacock SJ Prescott HC Pathophysiology transmission diagnosis and treatment of coronavirus disease 2019 COVID19 a review JAMA 2020 httpsdoiorg101001jama 202012839 2 McCulloch M Jezierski T Broffmann M Hubbard A Turner K Janecki T Diagnostic accuracy of canine scent detection in early and latestage lung and breast cancers Integr Cancer Ther 20065309 3 Guest C Pinder M Doggett M Squires C Affara M Kandeh B et al Trained dogs identify people with malaria parasites by their odour Lancet Infect Dis 20191957880 4 Taylor M McCready J Broukhanski G Kirpalaney S Lutz H Powis J Using dog scent detection as a pointofcare tool to identify toxigenic clostridium difficile in stool Open Forum Infect Dis 2018514 5 Angle C Waggoner LP Ferrando A Haney P Passler T Canine detection of the volatilome a review of implications for pathogen and disease detection Front Vet Sci 2016317 6 Angle TC Passler T Waggoner PL Fischer TD Rogers B Galik PK et al Real time detection of a virus using detection dogs Front Vet Sci 2016216 7 Koivusalo M Reeve C Biomedical scent detection dogs would they pass as a health technology Pet Behav Sci 2018617 8 Amann A Costello BDL Miekisch W Schubert J Buszewski B Pleil J et al The human volatilome volatile organic compounds VOCs in exhaled breath skin emanations urine feces and saliva J Breath Res 2014 https doiorg1010881752715583034001 9 Shirasu M Touhara K The scent of disease volatile organic compounds of the human body related to disease and disorder J Biochem 20111503 25766 10 Institut Pasteur Paris Protocol Realtime RTPCR assays for the detection of SARSCoV2 2020 Mar Available from httpswwwwhointdocsdefault Jendrny et al BMC Infectious Diseases 2020 20536 Page 6 of 7 sourcecoronaviruserealtimertpcrassaysforthedetectionofsarscov2 institutpasteurparispdfsfvrsn3662fcb62 Accessed 31 Mar 2020 11 Etievant S Bal A Escurret V BrengelPesce K Bouscambert M Cheynet V et al Sensitivity assessment of SARSCoV2 PCR assays developed by WHO referral Laboratories medRxiv 2020050320072207 httpsdoiorg1011012 020050320072207 12 HoffmannhttpswwwdvgnetfileadminBilderDVGPDF200331LA RT qPCRSARSCoV 2IP4AgPathpdf Accessed 31 Mar 2020 13 Hoffmann B Depner K Schirrmeier H Beer M A universal heterologous internal control system for duplex realtime RTPCR assays used in a detection system for pestiviruses J Virol Methods 20061362009 14 Sit THC Brackmann CJ Ip SM Tam KWS Law PYT To EMW et al Infection of dogs with ARSCoV2 Nature 2020 httpsdoiorg101038s4158602 023345 2020 15 Trevethan R Sensitivity specificity and predictive values foundations Pliabilities and pitfalls in research and practice Front Pub Health 20175 307 httpsdoiorg103389fpubh201700307 16 Cohen AN Kessel B 2020 False positives in reverse transcription PCR testing for SARSCoV2 medRxiv doi httpsdoiorg1011012020042620080911 17 Steppert C Steppert I Becher G Sterlacci W Bollinger T 2020 Rapid detection of SARSCoV2 infection by multicapillary column coupled ion mobility spectrometry MCCIMS of breath A proof of concept study medRxiv doihttpsdoiorg1011012020063020143347 18 Riley S Ainslie KEC Eales O Jeffrey B Walter CE Atchison C et al 2020 Community prevalence of SARSCoV2 virus in England during may 2020 REACT study medRxiv httpsdoiorg1011012020071020150524 19 Leroy EM Gouilh MA BrugèrePicoux J The risk of SARSCoV2 transmission to pets and other wild and domestic animals strongly mandates a one health strategy to control the COVID19 pandemic One Health 2020 httpsdoiorg101016jonehlt2020100133 20 Shi J Wen Z Zhong G Yang H Wang C Huang B et al Susceptibility of ferrets cats dogs and other domesticated animals to SARScoronavirus 2 Science 20203686494101620 Publishers Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations Jendrny et al BMC Infectious Diseases 2020 20536 Page 7 of 7