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Sampling and detection of airborne influenza virus towards point-of-care applications
KTH, School of Electrical Engineering (EES), Micro and Nanosystems.
KTH, School of Electrical Engineering (EES), Micro and Nanosystems.ORCID iD: 0000-0001-9177-1174
Janssen Diagnostics. (HMEEUWS1@its.jnj.com)
Janssen Diagnostics.
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2017 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, PlosONEArticle in journal (Refereed) Published
Abstract [en]

Airborne transmission of the influenza virus contributes significantly to the spread of this infectious pathogen, particularly over large distances when carried by aerosol droplets with long survival times. Efficient sampling of virus-loaded aerosol in combination with a low limit of detection of the collected virus could enable rapid and early detection of airborne influenza virus at the point-of-care setting. Here, we demonstrate a successful sampling and detection of airborne influenza virus using a system specifically developed for such applications. Our system consists of a custom-made electrostatic precipitation (ESP)-based bioaerosol sampler that is coupled with downstream quantitative polymerase chain reaction (qPCR) analysis. Aerosolized viruses are sampled directly into a miniaturized collector with liquid volume of 150 μL, which constitutes a simple and direct interface with subsequent biological assays. This approach reduces sample dilution by at least one order of magnitude when compared to other liquid-based aerosol bio-samplers. Performance of our ESP-based sampler was evaluated using influenza virus-loaded sub-micron aerosols generated from both cultured and clinical samples. Despite the miniaturized collection volume, we demonstrate a collection efficiency of at least 10% and sensitive detection of a minimum of 3721 RNA copies. Furthermore, we show that an improved extraction protocol can allow viral recovery of down to 303 RNA copies and a maximum sampler collection efficiency of 47%. A device with such a performance would reduce sampling times dramatically, from a few hours with current sampling methods down to a couple of minutes with our ESP-based bioaerosol sampler.

Place, publisher, year, edition, pages
Plos One , 2017.
National Category
Diagnostic Biotechnology Medical Laboratory and Measurements Technologies
Identifiers
URN: urn:nbn:se:kth:diva-199971DOI: 10.1371/journal.pone.0174314.s007ISI: 000399174400024Scopus ID: 2-s2.0-85016328510OAI: oai:DiVA.org:kth-199971DiVA, id: diva2:1067225
Note

QC 20170125

Available from: 2017-01-20 Created: 2017-01-20 Last updated: 2018-04-29Bibliographically approved
In thesis
1. An electrostatic sampling device for point-of-care detection of bioaerosols
Open this publication in new window or tab >>An electrostatic sampling device for point-of-care detection of bioaerosols
2018 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Bioaerosols are not only a significant factor of air quality but contribute greatly to the spread of infectious diseases, specifically through expired pathogen-laden aerosols. Clear examples of airborne transmission include: the recent influenza pandemic of 2009, the ongoing tuberculosis epidemic, and yearly norovirus out- breaks, which affect millions of people worldwide and pose serious threats to public healthcare systems. Given these acute concerns and the critical lack of knowledge of the field, it is important to develop methods for sampling and detecting these air- borne pathogens. Specifically, detection at the point-of-care can play an important role in improving the outcome of patient care by providing rapid and convenient diagnostics.

Electrostatic precipitation has emerged as a promising sampling tool for bio- aerosols, which together with a rapid analysis technique, can provide a powerful and integrated approach to pathogen detection or disease diagnosis at the point- of-care. Moreover, such a sampling-detection scheme could be a potentialy non- invasive breath sampling tool for diagnosis of respiratory infectious diseases.

This thesis presents a sampling device based on electrostatic precipitation, for capture of bioaerosols, and designed for use at point-of-care settings. A multi-point- to-plane electrode configuration allows charging of aerosol particles and direct air- to-liquid capture within a miniaturized volume with potentential for concatenation with on-site detection methods. Performance of the device was evaluated, using non-biological aerosols, for geometric (inter-electrode distance), electrical (inter- electrode potential and corona current), and aerosol parameters (particle size and gas velocity). Moreover, four different collector designs were investigated for im- proved collection efficiency and other features suitable for point-of-care settings (e.g. easy sample extraction and minimized volume).

The device was then validated, using bioaerosols, both in vitro and in vivo. In vitro validation was performed by capturing aerosolized influenza virus and analyz- ing the device collection efficiency. Lastly, prototype devices, designed for point- of-care, were validated in vivo with patients at the clinical setting. A pilot study was performed to capture exhaled pathogens from infected patients, with success- ful capture of exhaled bacteria.

Abstract [sv]

Bioaerosoler är inte enbart en signifikant faktor för luftkvalitet utan bidrar även mycket till spridningen av infektionssjukdomar, särskilt genom utandning av pato- genbelastade aerosoler. Tydliga exempel på luftburen överföring inkluderar: den se- naste influensapandemin under 2009, den pågående tuberkulosepidemin, och det årliga norovirusutbrott som påverkar miljontals människor världen över och utgör alvarliga hot mot folkhälsovårdssystemen. Med tanke på dessa akuta problem och den kritiska bristen på kunskap inom området är det viktigt att utveckla metoder för provtagning och detektering av dessa luftburna patogener. Specifikt kan detek- tion vid vårdpunkten spela en viktig roll för att förbättra resultatet as patientvården genom att tillhandahålla snabb och lämplig diagnostik.

Elektrostatisk utfällning har framkommit som ett lovande provtakningsverktyg för bioareosoler och kan, tillsammans med en snabb analys teknik, ge ett kraftfullt och integrerat tillvägagångsätt för detektering av patogener eller diagnos av sjuk- domar direkt vid vårdpunkten. Dessutom skulle ett sådant provtagning-detektions system kunna vara ett potentiellt icke-invasivt andprovtagningsverktyg för att dia- gnostisera infektionssjukdomar i andningsorganen.

Denna avhandling presenterar en elektrostatisk utfällningsenhet (ESP) för prov- tagning av bioaerosoler. Enheten är konstruerade för användning direkt på vård- platsen, har funktionen att fånga luft i vätska samt potential för sammanlänkning med platsens diagnosmetoder. En konfiguration med flera punkter-till-plan elektro- der möjliggör för laddning och avlägsnande av aerosolpartiklar för infångning i en vätskevolym på några hundra mikroliter. Infångning direkt i vätska ger inte enbart ett stabilt uppsamlings-, lagrings-, och transportmedium för patogener; det är även idealt för de flesta biologiska analyser, och den minimerade uppsamlingsvolymen resulterar i minimerade utspädningsfaktorer.

Utvecklingen av enheten beskrivs i den första delen, där icke-biologiska för- söksaerosoler används för att studera effekten av geometriska-, elektriska-, och ae- rosolparametrar på prestandan. Därutöver undersöks och optimeras utformningen av vätskekollektorn för elektrostatiskinfångning och hantering.

Validering av enheten, både in vitro och in vivo, beskrivs i den andra delen av av- handlingen och inkluderar användning biologiska aerosoler. Specifikt används ae- rosoliserad influensa för att genomföra en in vitro validering av ESP enheten, vars prestanda utvärderas för odlade och kliniska stammar. Slutligen utförs en validering in vivo vid vårdpunkten genom att använda ESP-enheten för att fånga patogener från utandningsluft.

Place, publisher, year, edition, pages
KTH Royal Institute of Technology, 2018. p. 45
Series
TRITA-EECS-AVL ; 2018:32
Keyword
electrostatic sampling; bioaerosols; point-of-care; breath sampling; electrostatic precipitation; microfluidic collection; corona discharge; air sampling; non-invasive diagnostics; exhaled breath analysis; on-site detection; aerosol sampling
National Category
Electrical Engineering, Electronic Engineering, Information Engineering Diagnostic Biotechnology
Research subject
Electrical Engineering
Identifiers
urn:nbn:se:kth:diva-226955 (URN)978-91-7729-748-2 (ISBN)
Public defence
2018-05-29, D3, Lindstedsvägen 5, Stockholm, 13:30 (English)
Opponent
Supervisors
Note

QC 20180502

Available from: 2018-05-02 Created: 2018-04-29 Last updated: 2018-05-02Bibliographically approved

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