On Chip Detection of Zika Virus Based on Loop Mediated Isothermal Amplification

Hong Yin; Xing Chen; Yong Jin; Bo Liu; Rongmeng Jiang

doi:doi:10.11648/j.ajbls.20210906.16

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On Chip Detection of Zika Virus Based on Loop Mediated Isothermal Amplification

Hong Yin, Xing Chen, Yong Jin, Bo Liu, Rongmeng Jiang

Published in American Journal of Biomedical and Life Sciences (Volume 9, Issue 6)

Received: 22 October 2021 Accepted: 19 November 2021 Published: 24 November 2021

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Abstract

There was a world-wide Zika virus (ZIKV) epidemic occurred in 2015, with the major concern of about 20-fold increase in fetuse microcephaly rate in Brazil. To improve the ZIKV point-of-care (POC) molecular diagnostic, we established a rapid and sensitive real-time fluorescence quantitative loop mediated isothermal amplification (LAMP) method and further applied it on a self-fabricated microfluidic chip. After optimization of LAMP reaction conditions, the assay achieved the detection limit of single copy of the standard plasmid in a reaction. Linear regression analysis revealed that the correlation coefficients (R²) were 0.9931. No cross reaction was observed in the controls of yellow fever clinical specimen and several known human influenza viruses, including seasonal A/H1N1, A/H7N9, A/H9N2 and B. To evaluate the performance characteristics of the ZIKV-LAMP assay, we detected 39 clinical specimens by both LAMP assay and real-time RT-PCR, which obtained with completely consistent results. The sensitivity, specificity and performance characteristics of the ZIKV-LAMP assay conformed its utility in ZIKV determination. Moreover, we had also developed a real-time fluorescence detection biomedical system with microfluidic chips. The microfluidic chips were designed with four microcells and the volume of the LAMP reaction was greatly reduced from about 25μL to 10μL. Our newly established real-time fluorescence LAMP detection system with microfluidic chips has the potential for ZIKV POC diagnostics with the advantages of low cost, short analytical time, disposability, low reagent and sample consumption and so on.

Published in	American Journal of Biomedical and Life Sciences (Volume 9, Issue 6)
DOI	10.11648/j.ajbls.20210906.16
Page(s)	302-306
Creative Commons	This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2021. Published by Science Publishing Group

Keywords

ZikaVirus, Fetal Microcephaly, Loop Mediated Isothermal Amplification Method, Microfluidic Chip

References

[1]	Mlakar, J. et al. Zika virus associated with microcephaly. N Engl J Med374, 951-958 (2016).
[2]	Rubin, E. J., Greene, M. F. & Baden, L. R. Zika virus and microcephaly. N Engl J Med374, 984-985 (2016).
[3]	Pardee, K. et al. Rapid, Low-Cost Detection of Zika Virus Using Programmable Biomolecular Components. Cell165, 1255-1266 (2016).
[4]	Faria, N. R. et al. Zika virus in the Americas: Early epidemiological and genetic findings. Science352, 345-349 (2016).
[5]	Dai, L. et al. Structures of the ZikaVirusEnvelope Protein and Its Complexwith a Flavivirus Broadly Protective Antibody. Cell Host & Microbe 19, 696–704 (2016).
[6]	Artron One Step Zika Virus Test Kit. (2016).
[7]	BiocanZika Rapid Test Approved in Brazil., (2016) (22/02/2016).
[8]	Charrel, R. N. et al. State of knowledge on Zika virus for anadequate laboratory. , (2016).
[9]	Faye, O. et al. Quantitative real-time PCR detection of Zika virus and evaluation with field-caught mosquitoes. Virol J10, 311 (2013).
[10]	Faye, O. et al. One-step RT-PCR for detection of Zika virus. J Clin Virol43, 96-101 (2008).
[11]	WHO, Current Zika Product Pipeline., (2016) (03/03/2016).
[12]	Notomi, T. et al. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 28, E63 (2000).
[13]	Hong, T. C. et al. Development and evaluation of a novel loop-mediated isothermal amplification method for rapid detection of severe acute respiratory syndrome coronavirus. J Clin Microbiol 42, 1956-1961 (2004).
[14]	Imai, M. et al. Development of H5-RT-LAMP (loop-mediated isothermal amplification) system for rapid diagnosis of H5 avian influenza virus infection. Vaccine24, 6679-6682 (2006).
[15]	Curtis, K. A., Rudolph, D. L. & Owen, S. M. Rapid detection of HIV-1 by reverse-transcription, loop-mediated isothermal amplification (RT-LAMP). J Virol Methods151, 264-270 (2008).
[16]	Wang, X. et al. Detection of enterovirus 71 using reverse transcription loop-mediated isothermal amplification (RT-LAMP). J Virol Methods179, 330-4 (2012).
[17]	Ge, Y. et al. Rapid and Sensitive Detection of Novel Avian-Origin Influenza A (H7N9) Virus by Reverse Transcription Loop-Mediated Isothermal Amplification Combined with a Lateral-Flow Device. PLoS One8, e69941 (2013).
[18]	Whitesides, G. M. The origins and the future of microfluidics. Nature442, 368-373 (2006).
[19]	Arora, A., Simone, G., Salieb-Beugelaar, G. B., Kim, J. T. &Manz, A. Latest developments in micro total analysis systems. Anal Chem 82, 4830-4847 (2010).
[20]	Stone, H. A., Stroock, A. D. & Ajdari, A. A. Engineering flows in smalldevices: microfluidics toward a Lab-on-a-Chip. Annu Rev Fluid Mech36, 381-411 (2004).
[21]	Herr, A. E. et al. Microfluidic immunoassays as rapidsaliva-based clinical diagnostics. ProcNatlAcadSci U S A104, 5268-5273 (2007).
[22]	Ferguson, B. S. et al. Genetic analysis of H1N1 influenza virus from throat swabsamples in a microfluidic system for point-of-care diagnostics. J AmChemSoc 133, 9129-9135 (2011).
[23]	Lingam, S., Beta, M., Dendukuri, D. &Krishnakumar, S. A. focus onmicrofluidics and nanotechnology approaches for the ultra sensitive detection of microRNAMicrorna 3, 18-28 (2014).
[24]	Uddin, S. M. et al. A Portable AutomaticEndpoint Detection System for Amplicons of Loop Mediated IsothermalAmplification on Microfluidic Compact Disk Platform. Sensors15, 5376-5389 (2015).
[25]	Luo, J. et al. A real-time microfluidic multiplex electrochemical loop-mediated isothermal amplification chip for differentiating bacteria. Biosens Bioelectron 60, 84-91 (2014).
[26]	Fang, X., Liu, Y., Kong, J. & Jiang, X. Loop-Mediated Isothermal Amplification Integrated on Microfluidic Chips for Point-of-Care Quantitative Detection of Pathogens. Anal Chem 82, 3002-3006 (2010).
[27]	Song, J. et al. Instrument-Free Point-of-Care Molecular Detection of Zika Virus. Anal Chem 88, 7289-7294 (2016).
[28]	Tian, B. et al. AttomolarZika virus oligonucleotide detection based on loop-mediated isothermal amplification and AC susceptometry. Biosens Bioelectron 86, 420-425 (2016).
[29]	Haddow, A. D. et al. Genetic Characterization of Zika Virus Strains: Geographic Expansion of the Asian Lineage. PLoS Negl Trop Dis. 6, e1477 (2012).

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APA Style

Hong Yin, Xing Chen, Yong Jin, Bo Liu, Rongmeng Jiang. (2021). On Chip Detection of Zika Virus Based on Loop Mediated Isothermal Amplification. American Journal of Biomedical and Life Sciences, 9(6), 302-306. https://doi.org/10.11648/j.ajbls.20210906.16

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ACS Style

Hong Yin; Xing Chen; Yong Jin; Bo Liu; Rongmeng Jiang. On Chip Detection of Zika Virus Based on Loop Mediated Isothermal Amplification. Am. J. Biomed. Life Sci. 2021, 9(6), 302-306. doi: 10.11648/j.ajbls.20210906.16

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AMA Style

Hong Yin, Xing Chen, Yong Jin, Bo Liu, Rongmeng Jiang. On Chip Detection of Zika Virus Based on Loop Mediated Isothermal Amplification. Am J Biomed Life Sci. 2021;9(6):302-306. doi: 10.11648/j.ajbls.20210906.16

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@article{10.11648/j.ajbls.20210906.16,
  author = {Hong Yin and Xing Chen and Yong Jin and Bo Liu and Rongmeng Jiang},
  title = {On Chip Detection of Zika Virus Based on Loop Mediated Isothermal Amplification},
  journal = {American Journal of Biomedical and Life Sciences},
  volume = {9},
  number = {6},
  pages = {302-306},
  doi = {10.11648/j.ajbls.20210906.16},
  url = {https://doi.org/10.11648/j.ajbls.20210906.16},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ajbls.20210906.16},
  abstract = {There was a world-wide Zika virus (ZIKV) epidemic occurred in 2015, with the major concern of about 20-fold increase in fetuse microcephaly rate in Brazil. To improve the ZIKV point-of-care (POC) molecular diagnostic, we established a rapid and sensitive real-time fluorescence quantitative loop mediated isothermal amplification (LAMP) method and further applied it on a self-fabricated microfluidic chip. After optimization of LAMP reaction conditions, the assay achieved the detection limit of single copy of the standard plasmid in a reaction. Linear regression analysis revealed that the correlation coefficients (R2) were 0.9931. No cross reaction was observed in the controls of yellow fever clinical specimen and several known human influenza viruses, including seasonal A/H1N1, A/H7N9, A/H9N2 and B. To evaluate the performance characteristics of the ZIKV-LAMP assay, we detected 39 clinical specimens by both LAMP assay and real-time RT-PCR, which obtained with completely consistent results. The sensitivity, specificity and performance characteristics of the ZIKV-LAMP assay conformed its utility in ZIKV determination. Moreover, we had also developed a real-time fluorescence detection biomedical system with microfluidic chips. The microfluidic chips were designed with four microcells and the volume of the LAMP reaction was greatly reduced from about 25μL to 10μL. Our newly established real-time fluorescence LAMP detection system with microfluidic chips has the potential for ZIKV POC diagnostics with the advantages of low cost, short analytical time, disposability, low reagent and sample consumption and so on.},
 year = {2021}
}

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TY  - JOUR
T1  - On Chip Detection of Zika Virus Based on Loop Mediated Isothermal Amplification
AU  - Hong Yin
AU  - Xing Chen
AU  - Yong Jin
AU  - Bo Liu
AU  - Rongmeng Jiang
Y1  - 2021/11/24
PY  - 2021
N1  - https://doi.org/10.11648/j.ajbls.20210906.16
DO  - 10.11648/j.ajbls.20210906.16
T2  - American Journal of Biomedical and Life Sciences
JF  - American Journal of Biomedical and Life Sciences
JO  - American Journal of Biomedical and Life Sciences
SP  - 302
EP  - 306
PB  - Science Publishing Group
SN  - 2330-880X
UR  - https://doi.org/10.11648/j.ajbls.20210906.16
AB  - There was a world-wide Zika virus (ZIKV) epidemic occurred in 2015, with the major concern of about 20-fold increase in fetuse microcephaly rate in Brazil. To improve the ZIKV point-of-care (POC) molecular diagnostic, we established a rapid and sensitive real-time fluorescence quantitative loop mediated isothermal amplification (LAMP) method and further applied it on a self-fabricated microfluidic chip. After optimization of LAMP reaction conditions, the assay achieved the detection limit of single copy of the standard plasmid in a reaction. Linear regression analysis revealed that the correlation coefficients (R2) were 0.9931. No cross reaction was observed in the controls of yellow fever clinical specimen and several known human influenza viruses, including seasonal A/H1N1, A/H7N9, A/H9N2 and B. To evaluate the performance characteristics of the ZIKV-LAMP assay, we detected 39 clinical specimens by both LAMP assay and real-time RT-PCR, which obtained with completely consistent results. The sensitivity, specificity and performance characteristics of the ZIKV-LAMP assay conformed its utility in ZIKV determination. Moreover, we had also developed a real-time fluorescence detection biomedical system with microfluidic chips. The microfluidic chips were designed with four microcells and the volume of the LAMP reaction was greatly reduced from about 25μL to 10μL. Our newly established real-time fluorescence LAMP detection system with microfluidic chips has the potential for ZIKV POC diagnostics with the advantages of low cost, short analytical time, disposability, low reagent and sample consumption and so on.
VL  - 9
IS  - 6
ER  -

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Author Information

Hong Yin

Institute of Equipment and Technology, Chinese Academy of Inspection and Quarantine, Beijing, China
Xing Chen

State Key Laboratory of Transducer Tech., Institute of Electronics, Chinese Academy of Sciences, Beijing, China
Yong Jin

Institute of Equipment and Technology, Chinese Academy of Inspection and Quarantine, Beijing, China
Bo Liu

Institute of Equipment and Technology, Chinese Academy of Inspection and Quarantine, Beijing, China
Rongmeng Jiang

Clinical and Research Center of Infectious Diseases, Beijing Ditan Hospital Capital Medical University, Beijing, China

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APA Style

Hong Yin, Xing Chen, Yong Jin, Bo Liu, Rongmeng Jiang. (2021). On Chip Detection of Zika Virus Based on Loop Mediated Isothermal Amplification. American Journal of Biomedical and Life Sciences, 9(6), 302-306. https://doi.org/10.11648/j.ajbls.20210906.16

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ACS Style

Hong Yin; Xing Chen; Yong Jin; Bo Liu; Rongmeng Jiang. On Chip Detection of Zika Virus Based on Loop Mediated Isothermal Amplification. Am. J. Biomed. Life Sci. 2021, 9(6), 302-306. doi: 10.11648/j.ajbls.20210906.16

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AMA Style

Hong Yin, Xing Chen, Yong Jin, Bo Liu, Rongmeng Jiang. On Chip Detection of Zika Virus Based on Loop Mediated Isothermal Amplification. Am J Biomed Life Sci. 2021;9(6):302-306. doi: 10.11648/j.ajbls.20210906.16

Copy | Download

@article{10.11648/j.ajbls.20210906.16,
  author = {Hong Yin and Xing Chen and Yong Jin and Bo Liu and Rongmeng Jiang},
  title = {On Chip Detection of Zika Virus Based on Loop Mediated Isothermal Amplification},
  journal = {American Journal of Biomedical and Life Sciences},
  volume = {9},
  number = {6},
  pages = {302-306},
  doi = {10.11648/j.ajbls.20210906.16},
  url = {https://doi.org/10.11648/j.ajbls.20210906.16},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ajbls.20210906.16},
  abstract = {There was a world-wide Zika virus (ZIKV) epidemic occurred in 2015, with the major concern of about 20-fold increase in fetuse microcephaly rate in Brazil. To improve the ZIKV point-of-care (POC) molecular diagnostic, we established a rapid and sensitive real-time fluorescence quantitative loop mediated isothermal amplification (LAMP) method and further applied it on a self-fabricated microfluidic chip. After optimization of LAMP reaction conditions, the assay achieved the detection limit of single copy of the standard plasmid in a reaction. Linear regression analysis revealed that the correlation coefficients (R2) were 0.9931. No cross reaction was observed in the controls of yellow fever clinical specimen and several known human influenza viruses, including seasonal A/H1N1, A/H7N9, A/H9N2 and B. To evaluate the performance characteristics of the ZIKV-LAMP assay, we detected 39 clinical specimens by both LAMP assay and real-time RT-PCR, which obtained with completely consistent results. The sensitivity, specificity and performance characteristics of the ZIKV-LAMP assay conformed its utility in ZIKV determination. Moreover, we had also developed a real-time fluorescence detection biomedical system with microfluidic chips. The microfluidic chips were designed with four microcells and the volume of the LAMP reaction was greatly reduced from about 25μL to 10μL. Our newly established real-time fluorescence LAMP detection system with microfluidic chips has the potential for ZIKV POC diagnostics with the advantages of low cost, short analytical time, disposability, low reagent and sample consumption and so on.},
 year = {2021}
}

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TY  - JOUR
T1  - On Chip Detection of Zika Virus Based on Loop Mediated Isothermal Amplification
AU  - Hong Yin
AU  - Xing Chen
AU  - Yong Jin
AU  - Bo Liu
AU  - Rongmeng Jiang
Y1  - 2021/11/24
PY  - 2021
N1  - https://doi.org/10.11648/j.ajbls.20210906.16
DO  - 10.11648/j.ajbls.20210906.16
T2  - American Journal of Biomedical and Life Sciences
JF  - American Journal of Biomedical and Life Sciences
JO  - American Journal of Biomedical and Life Sciences
SP  - 302
EP  - 306
PB  - Science Publishing Group
SN  - 2330-880X
UR  - https://doi.org/10.11648/j.ajbls.20210906.16
AB  - There was a world-wide Zika virus (ZIKV) epidemic occurred in 2015, with the major concern of about 20-fold increase in fetuse microcephaly rate in Brazil. To improve the ZIKV point-of-care (POC) molecular diagnostic, we established a rapid and sensitive real-time fluorescence quantitative loop mediated isothermal amplification (LAMP) method and further applied it on a self-fabricated microfluidic chip. After optimization of LAMP reaction conditions, the assay achieved the detection limit of single copy of the standard plasmid in a reaction. Linear regression analysis revealed that the correlation coefficients (R2) were 0.9931. No cross reaction was observed in the controls of yellow fever clinical specimen and several known human influenza viruses, including seasonal A/H1N1, A/H7N9, A/H9N2 and B. To evaluate the performance characteristics of the ZIKV-LAMP assay, we detected 39 clinical specimens by both LAMP assay and real-time RT-PCR, which obtained with completely consistent results. The sensitivity, specificity and performance characteristics of the ZIKV-LAMP assay conformed its utility in ZIKV determination. Moreover, we had also developed a real-time fluorescence detection biomedical system with microfluidic chips. The microfluidic chips were designed with four microcells and the volume of the LAMP reaction was greatly reduced from about 25μL to 10μL. Our newly established real-time fluorescence LAMP detection system with microfluidic chips has the potential for ZIKV POC diagnostics with the advantages of low cost, short analytical time, disposability, low reagent and sample consumption and so on.
VL  - 9
IS  - 6
ER  -

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