Journal:2019 novel coronavirus disease (COVID-19): Paving the road for rapid detection and point-of-care diagnostics

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Full article title 2019 novel coronavirus disease (COVID-19): Paving the road for rapid detection and point-of-care diagnostics
Journal Micromachines
Author(s) Nguyen, Trieu; Bang, Dang Duong; Wolff, Anders
Author affiliation(s) Technical University of Denmark
Primary contact Email: awol at dtu dot dk
Year published 2020
Volume and issue 11(3)
Article # 306
DOI 10.3390/mi11030306
ISSN 2072-666X
Distribution license Creative Commons Attribution 4.0 International
Website https://www.mdpi.com/2072-666X/11/3/306/htm
Download https://www.mdpi.com/2072-666X/11/3/306/pdf (PDF)

Abstract

We believe a point-of-care (PoC) device for the rapid detection of the 2019 novel coronavirus (SARS-CoV-2) is crucial and urgently needed. With this perspective, we give suggestions regarding a potential candidate for the rapid detection of the coronavirus disease 2019 (COVID-19), as well as factors for the preparedness and response to the outbreak of COVID-19.

Keywords: COVID-19, Wuhan, 2019 novel coronavirus, point-of-care detection, SARS-CoV-2, loop-mediated isothermal amplification, LAMP assay, polymerase chain reaction, PCR

Introduction

On January 30, 2020, the World Health Organization (WHO) declared a global public health emergency[1] over the outbreak of a novel coronavirus, called the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) (previously "2019 novel coronavirus" or "2019-nCoV"), which originated in Wuhan City, in the Hubei Province of China. On February 11, WHO officially named the disease the coronavirus disease 2019 (COVID-19).[2] Human-to-human transmission (Figure 1) has been confirmed by WHO and by the Centers for Disease Control and Prevention (CDC) of the United States[3], with evidence of person-to-person transmission from three different cases outside China, namely in the U.S.[4], Germany[5], and Vietnam.[6]


Fig1 Nguyen Micromachines2020 11-3.png

Figure 1. Illustration of the transmission of various coronaviruses, including SARS-CoV-2.[7] Current studies have suggested that the intermediate carriers may be snakes[8] or pangolins[9], but according to WHO the real source is still unknown.[10][11].

COVID-19 has continuously spread to 104 countries; the number of confirmed infections reached 109,343 on March 9, 2020[12], and the death toll in China has overtaken the SARS epidemic of 2002–2003 and has risen to 3,100.[2] To slow down the spread of COVID-19, at least 50 million people in China have been placed under lockdown.[13] On March 8, 2020, Italy also undertook the same measures, with the northern part of the country getting placed under lockdown, affecting 16 million people.[14] The reproduction number R0 (i.e., the average number of secondary cases generated by a typical infectious individual) is estimated to be 2.68, and the doubling time is estimated to be 6.4 days.[15]

The difference in terminology between "coronavirus" and "SARS-Cov-2" is detailed in Table 1.

Table 1. Difference between the term "coronavirus" and "SARS-CoV-2"
Term Description
Coronavirus (CoV) A large and diverse family of enveloped, positive-stranded RNA viruses, with a ~26–32 kilobase genome.[16] The Coronaviridae cover a broad host range, infecting many mammalian and avian species, and induce upper respiratory, gastrointestinal, hepatic, and central nervous system diseases.[17] In the last few decades, coronaviruses have been shown to be capable of also infecting humans. The outbreak of severe acute respiratory syndrome (SARS) in 2003, and, more recently, Middle East respiratory syndrome (MERS) have proved the lethality of coroanviruses when they cross the species barrier and infect humans.[18]
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) A new zoonotic human coronavirus, which was reported and announced by the Chinese Center for Disease Control and Prevention (CCDC) on January 9, 2020.[19] This novel coronavirus can lead to coroanvirus disease 2019 (COVID-19) in humans. In spite of the fact that the initial infected cases have been associated with the Huanan South China Seafood Market, the source of SARS-CoV-2 is still unknown (Figure 1). On January 30, 2020, the WHO declared a global public health emergency regarding the outbreak of COVID-19. On March 11, 2020, WHO declared the outbreak of COVID-19 a pandemic.

The need for a rapid detection method and portable detection devices

The manifestation of the COVID-19 infection is highly nonspecific, including respiratory symptoms, fever, cough, dyspnea, and viral pneumonia.[20] Thus, diagnostic tests specific to this infection are urgently required to confirm suspected cases, screen patients, and conduct virus surveillance.

In this scenario, a point-of-care (PoC) device (i.e., a rapid, robust, and cost-efficient device that can be used onsite and in the field, and which does not necessarily require a trained technician to operate[21]) is crucial for the detection of COVID-19. Figure 2 shows the dramatic impact of early detection of infectious diseases in controlling an outbreak.[22][23][24]


Fig2 Nguyen Micromachines2020 11-3.png

Figure 2. The dramatic impact of the rapid detection of infectious diseases in controlling and preventing an outbreak (adapted from the works of Ke et al.[22], Isere et al.[23], and Jones et al.[24]).

Such a PoC device can be used in (but is not limited to) an emergency situation, such as the Diamond Princess cruise ship case. Recently, it was reported that the Diamond Princess cruise ship had been quarantined in Yokohama, Japan, due to a serious spreading of COVID-19 on the ship, with at least 454 infected cases out of 3,700 passengers and crew (reported by WHO[25], February 17, 2020). The detection of COVID-19 may not have been prompt enough as they did not have enough test kits to diagnose all the passengers on the ship in order to timely respond to the rapid spreading of the disease.[26]

The current standard molecular technique that is now being used to detect SARS-CoV-2 is real-time reverse transcription polymerase chain reaction (rRT-PCR). This protocol has been documented and available online on the WHO website since January 17, 2020.[27] The testing procedure includes: (i) specimen collection; (ii) packing (storage) and shipment of the clinical specimens; (iii) (good) communication with the laboratory and providing needed information; (iv) laboratory testing; and (v) reporting the results. This rRT-PCR technique requires sophisticated laboratory equipment that is often located at a central laboratory (Biosafety level 2 or above).[4][27][28] Sample transportation is inevitable. As a consequence, the time required to obtain the results can be up to two or three days. In the case of a public health emergency such as the COVID-19 outbreak, this time-consuming process of sample testing is not only extremely disadvantageous, but also dangerous since the virus needs to be contained.

Another concern of PCR testing is that commercial PCR-based methods are expensive, depend upon technical expertise, and the presence of viral RNA or DNA does not always reflect acute disease.[29][30][31] Furthermore, using PCR, codetection with other respiratory viruses is frequently encountered in coronaviruse (CoV) testing, and the contribution of positive CoV PCR results to disease severity is not always explicitly exhibited.[29][30][31] Furthermore, as of February 2, 2020 in the United States, as mentioned in the Interim Guidelines for Collecting, Handling, and Testing Clinical Specimens from Persons Under Investigation (PUIs) for 2019 Novel Coronavirus, diagnostic testing for COVID-19 can be conducted only at the CDC, though from February 4 onwards, COVID-19 tests can also be done at laboratories designated by the CDC. Likewise in China, where the outbreak is ongoing, samples had to be sent to Beijing for testing, as reported on January 31.[32] On February 4, China’s own CDC deployed a mobile biosafety laboratory to Wuhan in the Hubei Province to assist with the response.[33][34] On February 5, an emergency test laboratory (biosafety level 2) run by BGI Group (with global heartquarters in Shenzhen, China) was set up in Wuhan in the Hubei Province to assist the COVID-19 epidemic.[35]

A potential candidate for rapid detection of SARS-CoV-2: Loop-mediated isothermal amplification (LAMP) assays in PoC devices

In order to overcome the current time-consuming and laborious detection technique using RT-PCR, an alternative molecular amplification technique should be deployed. Loop-mediated isothermal amplification (LAMP) reaction is a novel nucleic acid amplification technique that amplifies DNA with high specificity, efficiency, and rapidity under isothermal conditions. This method uses a set of four specially designed primers, in combination with a DNA polymerase with strand displacement activity[36] to synthesize target DNA up to 109 copies in less than an hour at a constant temperature of 65 °C. The final products are stem-loop DNAs with multiple inverted repeats of the target, bearing structures with a cauliflower-like appearance. LAMP has high specificity and sensitivity and is simple to perform; hence, soon after its initial development, it became an enormously popular isothermal amplification method in molecular biology, with applications in pathogen detection. LAMP uses strand-displacement polymerases instead of heat denaturation to generate a single-stranded template. As such, it has the advantage of running at a constant temperature, simultaneously reducing the cumbersomeness of a thermocycler as well as the energy required. LAMP technology is proven to be more stable[37] and more sensitive[38] in detection compared to PCR. Other advantages of LAMP compared to those of PCR are shown in Table 2.

Table 2. Comparison between PCR and loop-mediated isothermal amplification (LAMP) reactions
PCR LAMP
Thermal cycling: Multiple heating and cooling cycles; hence, bulky and cumbersome Isothermal and continuous amplification: Smaller, simpler, and therefor portable
Always requires sample concentration and preparation: Time-consuming More flexible: For detection of viruses such as influenza[39] or human norovirus, LAMP assay offers one-step detection.[40] Sample preparation steps are simplified.
Multiple protocols: Complicated and requires a skilled technician Single protocol: Faster
Reaction hindered by inhibitors Reaction tolerates inhibitors and is more stable
Diagnostic sensitivity (95%) is currently reported as lower than LAMP[38][40][41] Diagnostic sensitivity > 95%
Technique is established Technique is still being explored

We believe that the LAMP assay could be a potential candidate for the point-of-care device application in the detection of COVID-19. The Veterinary Validation of Point-of-Care Diagnostic Instrument (VIVALDI) project[42] is an example of using LAMP in a point-of-care device for the detection of a zoonotic virus causing respiratory symptoms (such as the strains responsible for Avian influenza). With PoC devices such as the VETPOD (Veterinary, Portable, Onsite Detection), which the VIVALDI project is validating[43], detection time can be less than one hour. Besides PoC devices using disposable polymer chips and LAMP assays, as in the VETPOD, a lateral flow strip (LFS) would also be a suitable candidate for the rapid and on-site detection of COVID-19. A device such as COVID-19 IgM/IgG Rapid Test of BioMedomics is a good example.[44] The sensitivity of the COVID-19 IgM/IgG Rapid Test is 88.66%, which is expected to be lower than the sensitivity of tests based on LAMP-reaction assays (>95%). Therefore, a combination of LFS and LAMP into one device could be an excellent candidate for PoC testing of COVID-19.

Other important factors in fighting COVID-19

Furthermore, alongside detecting and containing the virus, for the sake of a public health response regarding the dynamics of the outbreak, the socio-economic impact of COVID-19 is equally in urgent need. WHO announced that to fight the further spread of COVID-19, the international community had launched a U.S. $675 million preparedness and response plan to run from February through April 2020.[45] In 2003, the SARS-CoV virus pulled the world’s output down by $50 billion. The early estimation for the cost to the global economy as a result of the outbreak of COVID-19 is about $360 billion.[46] This is because China’s GDP shares were approximately 17% globally as of 2019, which was about four times higher than in 2003, and the confirmed infected cases (at the time of doing the economic estimation, i.e., at the beginning of February 2020) are more than two times larger than the total of SARS. Given that the number of infected cases (109,343 confirmed cases) is currently approximately 14 times larger than SARS cases[47], and that the death toll due to COVID-19 has surpassed that of the SARS epidemic, the economic impact of COVID-19 might be much larger than $360 billion.

Furthermore, in order to win the battle against this outbreak, information on the epidemiological characteristics, such as the identification of the animal reservoirs[48] (Figure 1) and the risk factor of the disease, is also essential. The intermediate host carrying the disease is important to identify not only for the current epidemic, but also to eliminate a future outbreak. Together with all the aforementioned factors, the race for a vaccination against COVID-19 is equally essential. Although at this stage there is no registered treatment or vaccine for COVID-19, Zhang has recently mentioned some potential interventions[49], such as nutritional interventions (Vitamin A, B, C, D, E, and other trace minerals such as zinc and iron). Due to the high percentage of identicality in the sequence (up to 82% of the genome structure) between SARS-COV-2 and the SARS-CoV virus, immuno-enhancers and other specific treatment that have been applied for SARS could also be considered[49] for treatment of SARS-CoV-2.

Acknowledgements

Author contributions

Conceptualization, T.N.; writing—original draft preparation, T.N.; manuscript revision, T.N, D.D.B., and A.W.; funding acquisition, D.D.B., A.W. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by the European Union’s Horizon 2020 research and innovation program, the CORONADX project, grant agreement No: 101003562, and the VIVALDI project, grant agreement No: 773422.

Conflicts of interest

The authors declare no conflict of interest.

References

  1. World Health Organization (31 January 2020). "Novel Coronavirus (2019-nCoV): Situation Report - 11" (PDF). World Health Organization. https://www.who.int/docs/default-source/coronaviruse/situation-reports/20200131-sitrep-11-ncov.pdf. Retrieved 13 March 2020. 
  2. 2.0 2.1 World Health Organization (8 March 2020). "Coronavirus disease 2019 (COVID-19): Situation Report - 48" (PDF). World Health Organization. https://www.who.int/docs/default-source/coronaviruse/situation-reports/20200308-sitrep-48-covid-19.pdf. Retrieved 13 March 2020. 
  3. Centers for Disease Control and Prevention (March 2020). "How COVID-19 Spreads". Centers for Disease Control and Prevention. https://www.cdc.gov/coronavirus/2019-ncov/prevent-getting-sick/how-covid-spreads.html?CDC_AA_refVal=https%3A%2F%2Fwww.cdc.gov%2Fcoronavirus%2F2019-ncov%2Fprepare%2Ftransmission.html. Retrieved 13 March 2020. 
  4. 4.0 4.1 Holshue, M.L.; DeBolt, C.; Lindquist, S. et al. (2020). "First Case of 2019 Novel Coronavirus in the United States". New England Journal of Medicine 382 (10): 929–36. doi:10.1056/NEJMoa2001191. PMC PMC7092802. PMID 32004427. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7092802. 
  5. Rothe, C.; Schunk, M.; Bretzel, G. et al. (2020). "Transmission of 2019-nCoV Infection from an Asymptomatic Contact in Germany". New England Journal of Medicine 382 (10): 970–71. doi:10.1056/NEJMc2001468. PMID 32003551. 
  6. Phan, L.T.; Nguyen, T.V.; Luong, Q.C. et al. (2020). "Importation and Human-to-Human Transmission of a Novel Coronavirus in Vietnam". New England Journal of Medicine 382 (9): 872-874. doi:10.1056/NEJMc2001272. PMID 31991079. 
  7. Enserink, M. (12 February 2020). "Update: ‘A bit chaotic.’ Christening of new coronavirus and its disease name create confusion". Science. https://www.sciencemag.org/news/2020/02/bit-chaotic-christening-new-coronavirus-and-its-disease-name-create-confusion. Retrieved 13 March 2020. 
  8. Ji, W.; Wang, W.; Zhao, X. et al. (2020). "Cross-species transmission of the newly identified coronavirus 2019-nCoV". Journal of Medical Virology 92 (4): 433-440. doi:10.1002/jmv.25682. PMID 31967321. 
  9. Cyrankoski, D. (7 February 2020). "Did pangolins spread the China caronavirus to people?". Nature - News. doi:10.1038/d41586-020-00364-2. https://www.nature.com/articles/d41586-020-00364-2. 
  10. World Health Organization (March 2020). "WHO recommendations to reduce risk of transmission of emerging pathogens from animals to humans in live animal markets or animal product markets". World Health Organization. https://www.who.int/health-topics/coronavirus/who-recommendations-to-reduce-risk-of-transmission-of-emerging-pathogens-from-animals-to-humans-in-live-animal-markets. Retrieved 13 March 2020. 
  11. Wang, W.; Tang, J.; Wei, F. (2020). "Updated understanding of the outbreak of 2019 novel coronavirus (2019‐nCoV) in Wuhan, China". Journal of Medical Virology 92 (4): 441–47. doi:10.1002/jmv.25689. 
  12. World Health Organization (March 2020). "Coronavirus disease (COVID-2019) situation reports". World Health Organization. https://www.who.int/emergencies/diseases/novel-coronavirus-2019/situation-reports/. Retrieved 09 March 2020. 
  13. Cohen, J.; Kupferschmidt, K. (2020). "Strategies shift as coronavirus pandemic looms". Science 367 (6481): 962–63. doi:10.1126/science.367.6481.962. PMID 32108093. 
  14. Regan, H. (8 March 2020). "Italy announces lockdown as global coronavirus cases surpass 105,000". CNN. https://edition.cnn.com/2020/03/08/asia/coronavirus-covid-19-update-intl-hnk/index.html. Retrieved 08 March 2020. 
  15. Wu, J.T.; Leung, K.; Leung, G.M. (2020). "Nowcasting and forecasting the potential domestic and international spread of the 2019-nCoV outbreak originating in Wuhan, China: A modelling study". Lancet 395 (10225): 689-697. doi:10.1016/S0140-6736(20)30260-9. PMID 32014114. 
  16. Tang, B.; Bragazzi, N.L.; Li, Q. et al. (2020). "An updated estimation of the risk of transmission of the novel coronavirus (2019-nCov)". Infectious Disease Modelling 5: 248-255. doi:10.1016/j.idm.2020.02.001. PMC PMC7029158. PMID 32099934. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7029158. 
  17. Gallagher, T.M.; Buchmeier, M.J. (2001). "Coronavirus spike proteins in viral entry and pathogenesis". Virology 279 (2): 371–4. doi:10.1006/viro.2000.0757. PMID 11162792. 
  18. Schoeman, D.; Fielding, B.C. (2019). "Coronavirus envelope protein: Current knowledge". Virology Journal 16 (1): 69. doi:10.1186/s12985-019-1182-0. PMC PMC6537279. PMID 31133031. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6537279. 
  19. Gralinski, L.E.; Menechary, V.D. (2020). "Return of the Coronavirus: 2019-nCoV". Viruses 12 (2): E135. doi:10.3390/v12020135. PMC PMC7077245. PMID 31991541. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7077245. 
  20. Huang, C.; Wang, T.; Li, X. et al. (2020). "Clinical features of patients infected with 2019 novel coronavirus in Wuhan, China". Lancet 395 (10223): 497–506. doi:10.1016/S0140-6736(20)30183-5. PMID 31986264. 
  21. Nguyen, T.; Zoëga Andreasen, S.; Wolff, A. et al. (2018). "From Lab on a Chip to Point of Care Devices: The Role of Open Source Microcontrollers". Micromachines 9 (8): E403. doi:10.3390/mi9080403. PMC PMC6187319. PMID 30424336. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6187319. 
  22. 22.0 22.1 Ke, C.-M.; Tsai, H.-C.; Chen, Y.-S. et al. (2015). "Outbreak investigation of pandemic influenza A H1N1 at the emergency department in a medical center in Southern Taiwan". Journal of Microbiology, Immunology and Infection 48 (2, Suppl. 1): S36. doi:10.1016/j.jmii.2015.02.051. 
  23. 23.0 23.1 Isere, E.E.; Fatiregun, A.A.; Ajayi, I.O. (2015). "An overview of disease surveillance and notification system in Nigeria and the roles of clinicians in disease outbreak prevention and control". Nigerian Medical Journal 56 (3): 161–8. doi:10.4103/0300-1652.160347. PMC PMC4518330. PMID 26229222. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4518330. 
  24. 24.0 24.1 Jones, G.; Le Hello, S.; Jourdan-da Silva, N. et al. (2014). "The French human Salmonella surveillance system: evaluation of timeliness of laboratory reporting and factors associated with delays, 2007 to 2011". Euro Surveillance 19 (1): 20664. doi:10.2807/1560-7917.es2014.19.1.20664. PMID 24434174. 
  25. World Health Organization (17 February 2020). "Coronavirus disease 2019 (COVID-19): Situation Report - 28" (PDF). World Health Organization. https://www.who.int/docs/default-source/coronaviruse/situation-reports/20200217-sitrep-28-covid-19.pdf. Retrieved 13 March 2020. 
  26. The Japan Times Staff (10 February 2020). "Coronavirus infection tally on Diamond Princess hits 135 as tests for all passengers eyed". The Japan Times. https://www.japantimes.co.jp/news/2020/02/10/national/japan-test-all-passengers-diamond-princess-cruise-ship-coronavirus/. Retrieved 10 February 2020. 
  27. 27.0 27.1 Corman, V.M.; Landt, O.; Kaiser, M. et al. (2020). "Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR". Euro Surveillance 25 (3): 2000045. doi:10.2807/1560-7917.ES.2020.25.3.2000045. PMC PMC6988269. PMID 31992387. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6988269. 
  28. Chu, D.K.W.; Pan, Y.; Cheng, S.M.S. et al. (2020). "Molecular Diagnosis of a Novel Coronavirus (2019-nCoV) Causing an Outbreak of Pneumonia". Clinical Chemistry 66 (4): 549-555. doi:10.1093/clinchem/hvaa029. PMC PMC7108203. PMID 32031583. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7108203. 
  29. 29.0 29.1 Bruning, A.H.L.; Aatola, H.; Toivola, H. et al. (2020). "Rapid detection and monitoring of human coronavirus infections". New Microbes and New Infections 24: 52–55. doi:10.1016/j.nmni.2018.04.007. PMC PMC5986163. PMID 29872531. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5986163. 
  30. 30.0 30.1 Gaunt, E.R.; Hardie, A.; Claas, E.C. et al. (2020). "Epidemiology and clinical presentations of the four human coronaviruses 229E, HKU1, NL63, and OC43 detected over three years using a novel multiplex real-time PCR method". Journal of Clinical Microbiology 48 (8): 2940-7. doi:10.1128/JCM.00636-10. PMC PMC2916580. PMID 20554810. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2916580. 
  31. 31.0 31.1 Cho, C.H.; Lee, C.K.; Nam, M.H. et al. (2014). "Evaluation of the AdvanSure real-time RT-PCR compared with culture and Seeplex RV15 for simultaneous detection of respiratory viruses". Diagnostic Microbiology and Infectious Disease 79 (1): 14–18. doi:10.1016/j.diagmicrobio.2014.01.016. PMID 24582583. 
  32. Brueck, H. (30 January 2020). "There's only one way to know if you have the coronavirus, and it involves machines full of spit and mucus". Business Insider. https://www.businessinsider.com/how-to-know-if-you-have-the-coronavirus-pcr-test-2020-1. Retrieved 13 March 2020. 
  33. Molteni, M. (4 February 2020). "The US Fast-Tracked a Coronavirus Test to Speed Up Diagnoses". Wired. https://www.wired.com/story/the-us-fast-tracked-a-coronavirus-test/. Retrieved 13 March 2020. 
  34. Chinese Center for Disease Control and Prevention (4 February 2020). "驰援武汉,高等级移动生物安全实验室启程". Chinese Center for Disease Control and Prevention. http://www.chinacdc.cn/yw_9324/202002/t20200204_212214.html. Retrieved 13 March 2020. 
  35. "New Emergency Detection Laboratory Run by BGI Starts Trial Operation in Wuhan, Designed to Test 10,000 Samples Daily". BGI News. BGI. 6 February 2020. https://www.bgi.com/global/company/news/new-emergency-detection-laboratory-run-by-bgi-starts-trial-operation-in-wuhan-designed-to-test-10000-samples-daily/. Retrieved 13 March 2020. 
  36. Nagamine, K.; Hase, T.; Notomi, T. (2002). "Accelerated reaction by loop-mediated isothermal amplification using loop primers". Molecular and Cellular Probes 16 (3): 223-9. doi:10.1006/mcpr.2002.0415. PMID 12144774. 
  37. Francois, P.; Tangomo, M.; Hibbs, J. et al. (2011). "Robustness of a loop-mediated isothermal amplification reaction for diagnostic applications". FEMS Immunology and Medical Microbiology 62 (1): 41–8. doi:10.1111/j.1574-695X.2011.00785.x. PMID 21276085. 
  38. 38.0 38.1 Galvez, L.C.; Barbosa, C.F.C.; Koh, R.B.L. et al. (2020). "Loop-mediated isothermal amplification (LAMP) assays for the detection of abaca bunchy top virus and banana bunchy top virus in abaca". Crop Protection 131: 105101. doi:10.1016/j.cropro.2020.105101. 
  39. Ahn, S.J.; Baek, Y.H.; Lloren, K.K.S. et al. (2019). "Rapid and simple colorimetric detection of multiple influenza viruses infecting humans using a reverse transcriptional loop-mediated isothermal amplification (RT-LAMP) diagnostic platform". BMC Infectious Diseases 19 (1): 676. doi:10.1186/s12879-019-4277-8. PMC PMC6669974. PMID 31370782. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6669974. 
  40. 40.0 40.1 Jeon, S.B.; Seo, D.J.; Oh, H. et al. (2017). "Development of one-step reverse transcription loop-mediated isothermal amplification for norovirus detection in oysters". Food Control 73 (Part B): 1002–9. doi:10.1016/j.foodcont.2016.10.005. 
  41. Wang, X.; Seo, D.J.; Lee, M.H. et al. (2014). "Comparison of conventional PCR, multiplex PCR, and loop-mediated isothermal amplification assays for rapid detection of Arcobacter species". Journal of Clinical Microbiology 52 (2): 557–63. doi:10.1128/JCM.02883-13. PMC PMC3911361. PMID 24478488. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3911361. 
  42. "VIVALDI". DTU Bioengineering, Technical University of Denmark. 2020. https://www.vivaldi-ia.eu/. Retrieved 13 March 2020. 
  43. "VIVALDI - Research". DTU Bioengineering, Technical University of Denmark. 2020. https://www.vivaldi-ia.eu/Research. Retrieved 13 March 2020. 
  44. "COVID-19 IgM/IgG Rapid Test". BioMedomics, Inc. 2020. https://www.biomedomics.com/products/infectious-disease/covid-19-rt/. Retrieved 13 March 2020. 
  45. World Health Organization (5 February 2020). "US$675 million needed for new coronavirus preparedness and response global plan". World Health Organization. https://www.who.int/news-room/detail/05-02-2020-us-675-million-needed-for-new-coronavirus-preparedness-and-response-global-plan. Retrieved 13 March 2020. 
  46. Raga, S. (5 February 2020). "Economic vulnerabilities to the coronavirus: Top countries at risk". ODI Blog. ODI. https://www.odi.org/blogs/16639-economic-vulnerabilities-coronavirus-top-countries-risk. Retrieved 13 March 2020. 
  47. Wang, C.; Horby, P.W.; Hayden, F.G. et al. (2020). "A novel coronavirus outbreak of global health concern". Lancet 395 (10223): 470-473. doi:10.1016/S0140-6736(20)30185-9. PMID 31986257. 
  48. Zhou, P.; Yang, X.L.; Wang, X.G. et al. (2020). "A pneumonia outbreak associated with a new coronavirus of probable bat origin". Nature 579 (7798): 270–73. doi:10.1038/s41586-020-2012-7. PMC PMC7095418. PMID 32015507. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7095418. 
  49. 49.0 49.1 Zhang, L.; Liu, Y. (2020). "Potential interventions for novel coronavirus in China: A systematic review". Journal of Medical Virology 95 (2): 479-490. doi:10.1002/jmv.25707. PMID 32052466. 

Notes

This presentation is faithful to the original, with only a few minor changes to presentation. In some cases important information was missing from the references, and that information was added. The original article referenced a generic situation report page from the WHO for the Diamond Princess infection numbers; the specific report (#28) is referenced for this version. Similarly, a more specific citation was used in this version for the mentioned VETPOD system. In a few places, "SARS-CoV-2" replaced "COVID-19" when the authors intended to reference the virus but referenced the disease.