logo

SCIENCE CHINA Materials, Volume 61, Issue 11: 1454-1461(2018) https://doi.org/10.1007/s40843-018-9253-5

Multiplexing technology for in vitro diagnosis of pathogens: the key contribution of phosphorus dendrimers

More info
  • ReceivedFeb 14, 2018
  • AcceptedMar 17, 2018
  • PublishedApr 16, 2018

Abstract

After the microbiology based on Pasteur’s method and polymerase chain reaction (PCR), the diagnosis company named Dendris has proposed a third-generation of diagnosis enabling the search of a broad range of pathogens with strong sensitivity and specificity. This extraordinary profile was possible thanks to the use of phosphorus dendrimers for which various techniques of deposition on a given support were investigated and described and analyzed in this report.


Funded by

the National Research Agency(Agence,Nationale,pour,la,Recherche)

“BIOTECHNOLOGIES” program(ANR,2010,BIOT,004,06:,Project,INNODIAG,to,JMF)

by Region Midi Pyrénées(06001324,&,07006292)

by CNRS.


Acknowledgment

This work was supported by the National Research Agency (Agence Nationale pour la Recherche), “BIOTECHNOLOGIES” program (ANR 2010 BIOT 004 06: Project INNODIAG to JMF) and by Region Midi Pyrénées (06001324 & 07006292) to RF and JMF and by CNRS (JPM, AMC).


Interest statement

The authors declare no conflict of interest.


Contributions statement

Majoral JP, Caminade AM designed experiments linked to dendrimer technology, François JM and Fabre R conceived the DendriSchip® kit fabrication. Majoral JP wrote the paper which was improved and approved by all the other authors.


Author information

Jean-Pierre Majoral is Emeritus Director of Research, Exceptional Class at the CNRS in Toulouse. His research interest is focused on the design and the properties of macromolecules such as phosphorus dendrimers and hyperbranched polymers. Main efforts are directed at the use of dendrimers in medicinal chemistry, material sciences and catalysis. He is co-founder and scientific director of the start-up Dendris. He is a member of several Academies of Sciences worldwide , got a dozen of international awards, and is an author of over 635 publications, 7 books, 35 book chapters, and 45 patents (h index 65, over 15,700 citations).


References

[1] He Z. Microarrays: Current Technology, Innovations, and Applications. Poole: Caister Academic Press, 2014. Google Scholar

[2] Jo H, Lee S, Ban C. Highly sensitive and selective in vitro diagnostics based on DNA probes and aptamers. Biodesign, 2015, 3: 33–40. Google Scholar

[3] Joos B, Kuster H, Cone R. Covalent attachment of hybridizable oligonucleotides to glass supports. Anal Biochem, 1997, 247: 96-101 CrossRef PubMed Google Scholar

[4] Rogers YH, Jiang-Baucom P, Huang ZJ, et al. Immobilization of oligonucleotides onto a glass support via disulfide bonds: a method for preparation of DNA microarrays. Anal Biochem, 1999, 266: 23-30 CrossRef PubMed Google Scholar

[5] Donatin E, Drancourt M. DNA microarrays for the diagnosis of infectious diseases. Médecine Maladies Infectieuses, 2012, 42: 453-459 CrossRef PubMed Google Scholar

[6] Pillet S, Lardeux M, Dina J, et al. Comparative evaluation of six commercialized multiplex PCR kits for the diagnosis of respiratory infections. PLoS ONE, 2013, 8: e72174 CrossRef PubMed ADS Google Scholar

[7] Benters R, Niemeyer CM, Wöhrle D. Dendrimer-activated solid supports for nucleic acid and protein microarrays. ChemBioChem, 2001, 2: 686-694 CrossRef Google Scholar

[8] Tomalia DA, Naylor AM, Goddard WA. Starburst dendrimers: molecular-level control of size, shape, surface chemistry, topology, and flexibility from atoms to macroscopic matter. Angew Chem Int Ed Engl, 1990, 29: 138-175 CrossRef Google Scholar

[9] Park JW, Jung Y, Jung YH, Seo JS, Lee Y. Preparation of oligonucleotide arrays with high-density DNA deposition and high hybridization efficiency. Bull Korean Chem Soc, 2004, 25: 1667–1670. Google Scholar

[10] Benters R. DNA microarrays with PAMAM dendritic linker systems. Nucleic Acids Res, 2002, 30: 10e-10 CrossRef Google Scholar

[11] Ahmed S, Vepuri SB, Kalhapure RS, et al. Interactions of dendrimers with biological drug targets: reality or mystery---a gap in drug delivery and development research. Biomater Sci, 2016, 4: 1032-1050 CrossRef PubMed Google Scholar

[12] Svenson S. The dendrimer paradox---high medical expectations but poor clinical translation. Chem Soc Rev, 2015, 44: 4131-4144 CrossRef PubMed Google Scholar

[13] Launay N, Caminade AM, Majoral JP. Synthesis and reactivity of unusual phosphorus dendrimers. a useful divergent growth approach up to the seventh generation. J Am Chem Soc, 1995, 117: 3282-3283 CrossRef Google Scholar

[14] Slomkowski S, Miksa B, Chehimi MM, et al. Inorganic–organic systems with tailored properties controlled on molecular, macromolecular and microscopic level. Reactive Funct Polymers, 1999, 41: 45-57 CrossRef Google Scholar

[15] Launay N, Caminade AM, Majoral JP. Synthesis of bowl-shaped dendrimers from generation 1 to generation 8. J Organomet Chem, 1997, 529: 51-58 CrossRef Google Scholar

[16] Le Berre V. Dendrimeric coating of glass slides for sensitive DNA microarrays analysis. Nucleic Acids Res, 2003, 31: 88e-88 CrossRef Google Scholar

[17] Trévisiol E, Le Berre-Anton V, Leclaire J, et al. Dendrislides, dendrichips: a simple chemical functionalization of glass slides with phosphorus dendrimers as an effective means for the preparation of biochips. New J Chem, 2003, 27: 1713-1719 CrossRef Google Scholar

[18] Chaize B, Nguyen M, Ruysschaert T, et al. Microstructured liposome array. Bioconjugate Chem, 2006, 17: 245-247 CrossRef PubMed Google Scholar

[19] Nicu L, Guirardel M, Chambosse F, et al. Resonating piezoelectric membranes for microelectromechanically based bioassay: detection of streptavidin–gold nanoparticles interaction with biotinylated DNA. Sensor Actuat B-Chem, 2005, 110: 125-136 CrossRef Google Scholar

[20] Thibault C, Le Berre V, Casimirius S, et al. Direct microcontact printing of oligonucleotides for biochip applications.. J Nanobiotechnol, 2005, 3: 7 CrossRef PubMed Google Scholar

[21] Feng CL, Zhong X  , Steinhart M, et al. Graded-bandgap quantum- dot-modified nanotubes: a sensitive biosensor for enhanced detection of DNA hybridization. Adv Mater, 2007, 19: 1933-1936 CrossRef Google Scholar

[22] Feng CL, Zhong XH, Steinhart M, et al. Functional quantum-dot/dendrimer nanotubes for sensitive detection of DNA hybridization. Small, 2008, 4: 566-571 CrossRef PubMed Google Scholar

[23] Yu Y, Feng C, Caminade AM, et al. The detection of DNA hybridization on phosphorus dendrimer multilayer films by surface plasmon field enhanced-fluorescence spectroscopy. Langmuir, 2009, 25: 13680-13684 CrossRef PubMed Google Scholar

[24] Feng CL, Yin M, Zhang D, et al. Fluorescent core-shell star polymers based bioassays for ultrasensitive DNA detection by surface plasmon fluorescence spectroscopy. Macromol Rapid Commun, 2011, 32: 679-683 CrossRef PubMed Google Scholar

[25] Jauvert E, Dague E, Séverac M, et al. Probing single molecule interactions by AFM using bio-functionalized dendritips. Senss Actuators B-Chem, 2012, 168: 436-441 CrossRef Google Scholar

[26] Patel JB. 16S rRNA gene sequencing for bacterial pathogen identification in the clinical laboratory. Mol Diagnosis, 2001, 6: 313-321 CrossRef Google Scholar

  • Figure 1

    Grafting of aldehyde terminated phosphorus dendrimer on aminosilanized glass slide.

  • Figure 2

    Microcontact printing of DNA involving four steps: 1) inking of the stamp with oligonucleotide, 2) drying under nitrogen stream, 3) contact of the glass slide functionalized by dendrimers with the inked PDMS stamp, 4) transfer of oligonucleotide from the PDMS stamp to obtain the patterned slide.

  • Figure 3

    Graded-bandgap structures based on charged phosphorus dendrimers and quantum dots inside an ordered porous alumina membrane.

  • Figure 4

    Hybridization of Cy5 complementary target DNA with DNA immobilized on phosphorus dendrimer multilayers (structures of the dendrimers are shown in Fig. 3).

  • Figure 5

    Structure of the anionic perylene diimide labelled star polymer A and of the cationic phosphorus dendrimer B.

  • Figure 6

    Sensitivity up to 10−18 obtained from a bilayer formed with A and B (Fig. 5) and using surface plasmon enhanced fluorescence spectroscopy technique.

  • Figure 7

    The DNA microarrays technique developed by Dendris.

  • Figure 8

    The advantages of DendrisChip® compared to other potential competitors. Reprinted with permission from [14].

  • Figure 9

    β-tester prototype proposed by Dendris.

  • Figure 10

    Sensitivity of the DendrisChips® technology (sum of blue and green ovals) versus microbiological method (sum of yellow and green ovals). Hi: H. influenzae; Pa: P. aeruginosa; Mca: M. catharralis; Kpn: K. pneumoniae; Sau: S. aureus. Caution : in the case of Sau, only 10 out of the 31 samples positively identified by the microbiological culture were confirmed by BDmax PCR assays.

  • Table 1   Comparative analysis of criteria from the different technologies used in IVD

    Culture method

    Mass spectrometry

    PCR

    Dendrischip®

    Multiplex

    No

    No

    Yes

    Yes

    Time of response

    48 h

    24 h

    1–3 h

    5 ha 

    Non-cultivable species

    No

    No

    Yes

    Yes

    Automatization

    No

    Semi

    Yes

    Yes

    Man power cost

    High

    Medium

    Low

    Low

    Including PCR amplification

Copyright 2020 Science China Press Co., Ltd. 《中国科学》杂志社有限责任公司 版权所有

京ICP备18024590号-1