Design and Applications of Fluorogenic Nucleic Acid-Based Paper Biosensors

Shuang Yang; Xianpeng Yang; Baojun Wang; Lei Wang

doi:10.7536/PC201129

Progress in Chemistry ›› 2021, Vol. 33 ›› Issue (12) : 2309-2315 DOI: 10.7536/PC201129

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Design and Applications of Fluorogenic Nucleic Acid-Based Paper Biosensors

Shuang Yang ¹^,²
, Xianpeng Yang ²
, Baojun Wang ³^,⁴^,⁵
, Lei Wang ²^,^*

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Abstract

In recent years, paper-based biosensors have attracted increasing attention due to their low cost, ease of operation and disposal, biodegradability and low consumption of analytes. Among them, the paper-based fluorescent biosensors with functional nucleic acids as the recognition elements are of particular attraction. Their high sensitivity, instant response and real-time detection capabilities endow them with great potentials for applications in portable sensor devices. In addition, the paper-based cell-free protein synthesis platform, using nucleic acid as the recognition elements, can achieve specific detection of viruses, heavy metals and other targets by expressing the fluorescent proteins as the output reporter, which has good application prospects. Here we introduce the design of these fluorogenic nucleic acid-based paper biosensors, focusing on the integration methods of nucleic acid-based recognition elements and paper-based substrates. We also discuss the latest progress of their applications in different fields including clinical diagnosis, food contaminant detection and environmental pollutant detection as well as their advantages and limitations. Finally, the prospects and development directions of fluorogenic nucleic acid-based paper biosensors are presented, providing reference for research in related fields.

Contents

1 Introduction

2 Design

2.1 Physical adsorption

2.2 Covalent coupling

2.3 Entrapment immobilization

3 Applications

3.1 Clinical diagnosis

3.2 Food safety detection

3.3 Environmental pollutant detection

4 Conclusions and outlook

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Keywords

nucleic acids / functional nucleic acids / paper-based materials / fluorescent response / biosensing

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Shuang Yang, Xianpeng Yang, Baojun Wang, Lei Wang. Design and Applications of Fluorogenic Nucleic Acid-Based Paper Biosensors. Progress in Chemistry, 2021, 33(12): 2309-2315 DOI:10.7536/PC201129

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1 Introduction

In recent years, the research of Point-of-Care Testing (POCT) devices has promoted the development of clinical diagnosis, food safety testing and environmental pollutant detection^[1]. The traditional detection technology needs to be carried out in the laboratory, equipped with expensive experimental instruments and skilled staff. In contrast, POCT devices have the advantages of low cost, fast and efficient, easy to use, and show great potential in the field of portable sensing and detection, especially in developing countries and regions where resources are relatively scarce^[2⇓~4]. According to the World Health Organization, diagnostic equipment in developing countries should be affordable, sensitive, specific, user-friendly, fast and stable, equipment-free and widely accessible to end users^[5,6]. Paper has become one of the ideal materials for POCT devices because of its low cost, good biocompatibility, abundant sources and biodegradability^[7⇓~9].

Paper-based detection technology began in the 1950s. Up to now, pregnancy test strips and blood glucose test strips have been widely used as simple biosensing devices for instant diagnosis and disease detection^[10⇓~12]. Biosensing devices use biomolecules as recognition elements, capture the analyte to be detected through the specific reaction between biomolecules and target molecules, and convert the specific reaction into detectable optical, electrical, acoustic, thermal and other signals through signal conversion elements^[13]. The performance of paper-based biosensing devices depends on the characteristics of the biological recognition elements carried by them, and the key to the preparation of paper-based biosensing devices is how to fix the biological recognition elements on the paper-based materials^[14]. Paper-based biosensors with functional nucleic acids or nucleic acids as recognition elements have been extensively studied in the past few years. Functional nucleic acids (FNAS) are natural or artificial nucleic acid sequences with specific structure and function, which have the functions of recognition, transformation, catalysis, color development, luminescence, electron transfer and so on. Functional nucleic acids isolated from nucleic acid sequence libraries can recognize or respond to almost any target to be detected, which is also the characteristic and advantage of nucleic acid-based paper-based biosensors compared with other paper-based biosensors^[15]. Natural functional nucleic acids include Ribozymes and Riboswitches; Artificial functional nucleic acids include Aptamers, Ribozymes and DNAzymes^[16]. Due to its high stability and low cost, the current research mainly focuses on Aptamers and DNAzymes^[17]. Nucleic acids (NAS) is the general name of deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). Cell-free protein synthesis system (CFPS) using exogenous DNA or mRNA as template is independent of cells, which breaks through the limitations of traditional biotechnology and has the advantages of green, economic, simple and fast. In recent years, it has become a research hotspot^{[18⇓⇓⇓~22]}. Paper-based CFPS biosensing devices are developed from whole-cell biosensing devices, both of which have the disadvantage of slow response, but compared with the latter, the former has the advantage that it will not cause safety problems of genetically modified organisms^[23]. Paper-based sensors containing different identification elements have different preparation methods, and have different advantages and disadvantages according to different application fields.

At present, the signal transduction mechanisms of nucleic acid-based paper-based biosensors mainly include electrochemical method, colorimetric method and fluorescence method^[24]. The fluorescence method is to recognize and respond to the input signal through the recognition element, and to realize the sensing response to the target by the change of the intensity of the fluorescence response signal. Fluorescence method can be used to observe the change of fluorescence signal with the help of smart phones and other communication devices, which is simple to operate, fast to respond and highly sensitive, and has been widely developed and applied to the detection of markers in various fields. Fluorescence biosensors using functional nucleic acids as recognition elements usually induce fluorescence resonance energy transfer (FRET) according to the change of the distance between the fluorescent donor group and the fluorescent acceptor group, resulting in positive fluorescence enhancement or negative fluorescence reduction, thus realizing the sensing response to the target^[25]. Fluorescent biosensors with nucleic acids as recognition elements are often based on CFPS, which can produce different fluorescence intensities by synthesizing fluorescent proteins to achieve the sensing response to the target^[26,27].

In this paper, we focus on paper-based fluorescent biosensors based on nucleic acid. Firstly, we introduce three methods of binding paper-based materials with nucleic acid or functional nucleic acid: physical adsorption, covalent coupling and embedding immobilization. The current research progress of nucleic acid-based paper-based fluorescent biosensors in three key areas, including clinical diagnosis, food safety detection and environmental pollutant detection, was summarized. Finally, the future research and development directions of this kind of equipment in the preparation process and application fields are prospected.

2 Design of a Paper-based Fluorescent Biosensor Based on Nucleic Acid

Paper-based fluorescent biosensors based on nucleic acid are mainly composed of two parts, usually paper-based materials as the carrier, and nucleic acid or functional nucleic acid as the recognition element. In addition, for in vitro transcription-translation systems, CFPS (cell extracts, amino acids, enzymes, nucleotides, etc.) need to be added for nucleic acid expression and normal operation. When designing a paper-based fluorescent biosensor based on nucleic acid, it is first necessary to firmly bind the nucleic acid or functional nucleic acid to the paper-based material and maintain its functional activity. The commonly used preparation methods are summarized as follows.

2.1 Physical adsorption

Physical adsorption refers to the process in which the recognition element is attached to the paper surface through non-covalent interactions such as hydrogen bonding, electrostatic interaction, and van der Waals force. The schematic diagram of physical adsorption fixation is shown in Figure 1 (a)^[28]. The structure of paper, such as physical pore size, accessible surface area, cellulose crystallinity, etc., and the surface chemistry of paper, such as surface energy, surface sizing agents, surface plasma treatment, etc., will affect the effect of paper adsorption and fixation^[14]. The surface of cellulose is negatively charged, so cationic polymers are easily adsorbed on the surface of cellulose, but there are negatively charged phosphate groups in DNA molecules, which are not easily adsorbed^[29]. Su et al. Used physical adsorption method to combine cellulose with DNA aptamer, and found that the aptamer was easily washed off and could only stay on the fiber surface for a short time^[30]. The advantage of physical adsorption is simple, economical and fast, and it does not need to pretreat the paper, which can avoid the denaturation of the recognition element. However, this method has great limitations, including the randomness of the adsorption process, the movement of the identification element when it contacts with the target, and the easy detachment from the paper-based material. These shortcomings may lead to poor reproducibility and low accuracy of test results, which are not suitable for the detection of high-precision results.

2.2 Covalent coupling

Covalent coupling immobilization is a method of binding the recognition element and the paper-based material together through covalent bonds. Before using this method, the recognition element and the paper-based material need to be chemically modified. This method is an ideal method for uniform and stable binding of the recognition element with the paper-based material. The schematic diagram of covalent coupling immobilization is shown in Figure 1 (B)^[30]. Usually, functional nucleic acids as recognition elements are combined with paper-based materials by covalent coupling technology. The common method is to convert the hydroxyl groups of cellulose into functional groups with higher chemical activity, such as aldehyde groups, epoxy groups and sulfonates^[31,32]^[33]^[34]. Su et al. Immobilized adenosine triphosphate (ATP) -bound DNA aptamers on cellulose by physical adsorption and covalent coupling to detect pathogens, and compared them^[30]. The former is to add cellulose into a buffer solution with functional nucleic acid to realize physical adsorption; In the latter, the cellulose is modified by aldehyde group, and then reacted with the aptamer with terminal ammonia through Schiff base to realize covalent coupling. Compared with physical adsorption, covalent coupling makes functional nucleic acids on paper more stable and more evenly distributed. Therefore, the sensor based on covalent coupling is more suitable for the detection of liquid samples, and it is also more suitable for the harsh test environment, and the experimental results also have good reproducibility. However, covalent coupling requires activation or complex chemical modification of the recognition element and the paper-based material, which may lead to structural changes in the recognition element and partial loss of activity, resulting in decreased sensitivity of the biosensor^[28].

2.3 Embedding fixation

In order to overcome the limitations of physical adsorption and covalent coupling methods, scholars have developed embedding immobilization methods, mainly including gel embedding and freeze-drying. The schematic diagrams of the two immobilization methods are shown in Figure 1 (C) and (d) respectively. Gel embedding is to embed nucleic acid or functional nucleic acid in the polymer network of gel, which is usually composed of synthetic polymers such as polyacrylamide and calcium alginate or natural polymers such as gelatin and starch. Su et al. reported a method for embedding DNA aptamers in microgels^[35]. The method first combines streptavidin with carboxyl modified poly (N-isopropylacrylamide) (PNIPAM) microgels. The biotin-bound aptamers have a high affinity for streptavidin, and the aptamers can be easily immobilized in microgels to form DNA aptamer microgels, which can be printed on paper. The results showed that the DNA aptamer was stable in the microgel, the recognition ability remained intact, and the binding between the microgel and the paper-based material was stable. Freeze-drying is to freeze the solution of nucleic acid or functional nucleic acid into solid in advance, and then dehydrate and dry it under vacuum condition. Pardee et al. reported a method of embedding CFPS into paper by freeze-drying^[36]. Firstly, the CFPS solution was dropped onto the paper, and the CFPS solution was quickly fixed in the porous network of the paper, and then the water was quickly removed by vacuum freeze-drying. This method creates paper-based bioactive materials with basic transcription and translation properties of cells, providing cheap, sterile and non-living cell-based synthetic biotechnology for clinical, global health, industry, research and other fields^[37]. Compared with the living cell system, CFPS is more controllable and operable. For example, in the process of protein synthesis, the system can be artificially intervened at any time to regulate protein expression^[18]. However, the high cost of the freeze-drying process may be an obstacle to the expanded use of cell-free protein synthesis techniques. To this end, Karig et al. Used trehalose as a protective agent and replaced the freeze-drying technique with a natural drying technique^[38]. The results showed that the reaction system was still active after being stored at 37 ℃ for several months, and could also be stored under atmospheric oxygen and humidity conditions.

3 Application of Paper-based Fluorescent Biosensor Based on Nucleic Acid

At present, three types of paper-based biosensors based on nucleic acid have been widely studied: electrochemical biosensors, colorimetric biosensors and fluorescent biosensors. They can be applied to a series of fields, such as clinical diagnosis, detection of environmental pollutants and detection of food risk factors^{[39⇓⇓⇓⇓~44]}^{[21,37,45⇓⇓ ~48]}^{[49⇓⇓⇓~53]}. Electrochemical biosensors (EBS) can achieve selective quantitative analysis by converting target molecules into electrochemically detectable substances through recognition elements, which has high sensitivity, but its preparation process involves many chemical reactions among the solid electrode surface, the recognition layer and the target in the solution, and the preparation and use process is relatively complex^[54⇓~56]. Colorimetric sensor is a sensing system that uses the color change of the system before and after the recognition reaction as the output signal^[57]. In contrast, colorimetric biosensors and fluorescent biosensors are easier to use, and the real-time response of fluorescence makes fluorescent biosensors have the advantage of real-time detection^[15]. Therefore, the nucleic acid-based paper-based fluorescent biosensing device is a promising sensing detection platform.

3.1 Clinical diagnosis

In the past few years, there has been a significant increase in the development of new paper-based biosensors, especially for the initial diagnosis of diseases^[58⇓~60]. Cheap paper-based sensing devices make disease diagnosis easy and fast. Takahashi et al. Developed a paper-based diagnostic platform based on synthetic biology, which was formed by embedding ribose Toehold switch sensors (TSS) and CFPS on a paper plate through freeze-drying.The system can perform low-cost, on-demand, and simple analysis of microflora samples. The workflow for analyzing biological samples is shown in Figure 2. When in contact with the target, the system can express Green fluorescent protein (GFP)^[61]. The researchers selected 10 disease-associated bacteria and designed the corresponding TSS to semi-quantitatively detect the specific mRNA of the bacteria. At the same time, they developed TSS for four human biomarkers, including calreticulin and tumor suppressor protein M, to analyze stool samples from patients with inflammatory bowel disease, and compared Reverse transcription quantitative PCR (RT-qPCR) technology. In addition, the platform can also be used for rapid detection of toxin mRNA to diagnose Clostridium infection (CDI), which is superior to the standard qPCR method. In the DNA-based qPCR test, all samples tested positive, resulting in test results that could not distinguish the degree of infection. When using paper-based diagnostic platform to detect toxin mRNA, we can quickly and easily distinguish CDI carriers expressing low levels of toxin from CDI patients expressing high levels of toxin. In addition to a wide range of biological targets such as viruses, fungi and target nucleic acids from fecal or tissue samples, this platform may also be used in the future to pre-screen participants in microbiome therapy trials and track changes in disease activity in patients.

3.2 Food safety testing

Food safety is closely related to human life and health, which is attracting the attention of all countries in the world. In the food industry, the quality control of food is very important before the product enters the market, that is, from the production stage to the packaging stage. Contamination of food with viruses, bacteria, parasites, heavy metal ions, pesticides and other adulterated chemicals can have serious consequences for human health and the economy^[62,63]. In the field of food safety, the matrix is complex, the residues of agricultural and veterinary drugs, the content of heavy metals and the amount of additives are small, and the detection methods need to have good selectivity, low detection limit and high sensitivity. These are the advantages of paper-based fluorescent biosensors based on nucleic acids^[64]. Weng et al. Used fluorescein (FAM) labeled norovirus specific aptamers to physically adsorb multi-walled carbon nanotubes (MWCNTs) and graphene oxide (GO), respectively.Immobilized on a paper substrate, two paper-based microfluidic devices were prepared to detect the possible presence of norovirus in complex food matrices. The design principle and the paper-based microfluidic devices are shown in 3 (B) and 3 (C), respectively^[53]. Among them, MWCNT and GO, as two different fluorescence quenchers, quench the fluorescence of FAM-labeled specific aptamer through FRET. Only in the presence of norovirus, the labeled aptamer specifically binds to the virus, and the aptamer is released from MWCNT or GO, and the fluorescence is recovered. The principle of fluorescence quenching-recovery is shown in Figure 3 (a). The detection limits of MWCNT and GO were 4.4 ng·mL^-1 and 3.3 ng·mL^-1, respectively. The development of this device provides a new method for early identification of norovirus and an intervention to prevent the spread of the epidemic, and is expected to be applied to the clinical diagnosis of norovirus infection in the future.

3.3 Detection of environmental pollutants

In recent years, there are more and more toxic and harmful pollutants in the environment, which have caused certain harm to the survival of human beings and animals, so the demand for the detection of environmental pollutants is also increasing^[65⇓~67]. For samples such as heavy metals and pesticide residues in water and soil, the process of collecting and transporting them back to the laboratory may lead to fluctuations in the sample environment, while testing is still largely limited to complex, centralized laboratories, which is time-consuming and requires a large number of technically specialized staff^[68⇓~70]. At present, researchers are also trying to explore and develop paper-based fluorescent biosensors with low cost and real-time detection of environmental pollutants. Khoshbin et al. developed a paper-based sensing device for detecting GGGT GGGT GGGT GGGT in lake water based on a DNA aptamer (5 '-FAM-GGGT GGGT GGGT-3') as a recognition element. The device has a wide range of applications and can also be used to detect GGGT GGGT GGGT GGGT in foods such as tap water and milk^[71]. The detection principle is shown in Figure 4. After the FAM-labeled aptamer is bound to the paper through simple physical adsorption, the aptamer is in a stretchable single-stranded state. When the GO solution is dropped onto the paper surface, π-π conjugation and hydrogen bonding interactions are formed between the nucleotide base of the aptamer and the aromatic structure of GO, which leads to the fluorescence quenching of the aptamer through FRET. When the sample containing Pb²⁺ was injected onto the GO-bearing paper surface, the conformation of the aptamer was changed from a random single strand to a G-quadruplet structure, the fluorophore was separated from the GO, and the fluorescence was restored again. The device is fast and sensitive, and can complete the detection within 10 min with a minimum detection limit of 0. 5 pM. In the future, other metal ions and biomolecules can also be effectively detected by replacing target-specific aptamers.

4 Conclusion and prospect

Nucleic acid-based biosensors have the advantages of fast response and strong specificity, and can identify or respond to more targets to be detected through functional modification or gene recombination technology. Paper is cheap, with porous network structure, biodegradable and good biocompatibility, which is suitable for the substrate of portable biosensors. Therefore, nucleic acid-based fluorescent biosensors based on paper combine the advantages of nucleic acids or functional nucleic acids and paper, and are widely used in clinical diagnosis, food safety testing, environmental testing and other fields, playing the role of simple, fast and real-time detection, which is an attractive POCT platform. At present, paper-based fluorescent biosensors based on nucleic acid have been widely studied, but further research and development are still needed to overcome some problems in the practical process.

(1) Design optimization or development of more efficient sensing and recognition elements. Compared with laboratory detection technology and clinical detection technology, POCT technology has developed rapidly in recent years, but the technology is not mature and the user trust is not enough. High sensitivity and specificity are still the primary indicators for paper-based fluorescent biosensors to be improved. Therefore, it is necessary to design and develop sensor recognition elements with excellent performance or further optimize the existing recognition elements with application potential to solve practical problems, such as early detection of cancer or other human diseases. In addition, as an in vitro life simulation system, the paper-based CFPS system is still in the early stage of research. With the rapid development of genetic engineering and synthetic biology, different novel and efficient recognition elements will be integrated into reporter genes simultaneously in the future, thus realizing the simultaneous detection of multiple analytes^[72].

(2) To develop a new method for the structural design of paper-based fluorescent biosensors. The combination of paper-based materials with nucleic acids or functional nucleic acids is not limited to physical adsorption, covalent coupling and embedding, such as the 3D printing technology which can integrate information, materials, biology, control and other technologies.It may provide a new way for the combination of paper-based materials and recognition elements, and realize the unification of paper-based fluorescent biosensors in structure, material and function through intelligent manufacturing. In addition, the functional modification of paper-based materials will also have a certain impact on the combination of paper-based materials and recognition elements.Compared with physical and chemical modification, the modified cellulose synthesized in situ by microorganisms provides a new idea for the functionalization of paper-based material^[73]. At the same time, the size of cellulose will also affect the pore size of paper-based materials, and then affect the binding of recognition elements and the reading of fluorescence signals.

(3) Intelligent monitoring and detection results. With the development of science and technology, paper-based fluorescent biosensors based on nucleic acid can be combined with smart phones, or through the development of various effective and portable intelligent signal readout devices, the detection results can be read anytime and anywhere, and gradually tend to be stable and universal, thereby reducing the dependence on professional detection equipment and professional technicians. At the same time, it will become a trend to develop paper-based fluorescent biosensors from simple qualitative analysis to semi-quantitative and complete quantitative analysis with the help of smart phones.

In addition, the application of paper-based fluorescent biosensors based on nucleic acid in different fields is still worthy of attention in future research. For the field of clinical diagnosis, it is particularly important to develop simple, fast and effective methods to process and detect real-life samples, such as feces or human serum, so POCT devices such as nucleic acid-based paper-based fluorescent biosensors can be truly used for clinical or personal use. In the field of food safety detection, the functional integration of paper-based fluorescent biosensors should be improved, such as the development of a multi-channel analysis system that can identify a variety of bacteria at one time, which will help to further reduce the analysis time and reduce the cost of detection. In the field of environmental pollutant detection, most of the existing paper-based fluorescent biosensors are still in the laboratory stage of detecting synthetic samples due to their low selectivity, and further development of paper-based fluorescent biosensors that can be applied to real environmental samples is needed. In a word, the future development of paper-based fluorescent biosensors based on nucleic acid will be simplified, portable, intelligent and practical, which will provide an opportunity to promote global POC detection and intelligent monitoring.

References

Publishing order | Descend order by publishing year | Descend order by cited within

[1]	Liana D D, Raguse B, Gooding J J, Chow E. Sensors, 2012, 12(9): 11505.

[2]	Zhang D H, Broyles D, Hunt E A, Dikici E, Daunert S, Deo S K. Anal., 2017, 142(5): 815.

[3]	Cinti S, Moscone D, Arduini F. Nat. Protoc., 2019, 14(8): 2437.

[4]	Wan X Y, Volpetti F, Petrova E, French C, Maerkl S J, Wang B J. Nat. Chem. Biol., 2019, 15(5): 540.

[5]	Peeling R W, Mabey D. Clin. Microbiol. Infect., 2010, 16(8): 1062.

[6]	Martinez A W, Phillips S T, Whitesides G M, Carrilho E. Anal. Chem., 2010, 82(1): 3.

[7]	Ge S G, Zhang L N, Yu J H. Bioanalysis, 2015, 7(6): 633.

[8]	Zhao W, Van D B A. Lab. Chip., 2008, 8(12): 1988.

[9]	Hui C Y, Liu M, Li Y F, Brennan J D. Angew. Chem. Int. Ed., 2018, 57(17): 4549.

[10]	Burstein J, Braunstein G D. Early pregnancy, 1995, 1(4): 288.

[11]	Chard T. Hum. Reprod., 1992, 7(5): 701.

[12]	Comer J P. Anal. Chem., 1956, 28(11): 1748.

[13]	Liu X F, Wang L H, Song S P, Fan C H, Huang W. Prog. Chem., 2008, 20(9): 1375.

[14]	( 刘兴奋, 王丽华, 宋世平, 樊春海, 黄维. 化学进展, 2008, 20(9): 1375.)

[15]	Kong F Z, Hu Y F. Anal. Bioanal. Chem., 2012, 403(1): 7.

[16]	Liu R D, McConnell E M, Li J X, Li Y F. J. Mater. Chem. B, 2020, 8(16): 3213.

[17]	Du Z H, Li X Y, Tian J J, Zhang Y Z, Tian H T, Xu W T. Chinese Journal of Analytical Chemistry, 2018, 46(7): 11.

[18]	( 杜再慧, 李相阳, 田晶晶, 张洋子, 田洪涛, 许文涛. 分析化学, 2018, 46(7): 11.)

[19]	Zhu L, Ling J J, Zhu Z, Tian T, Song Y L, Yang C Y. Anal. Bioanal. Chem., 2021, 413(18): 4563.

[20]	Liu Y, Guo X C, Geng J H, Jiao Y, Han J P, Zhang Z K, Zhou X, Yang D Y. Chinese Science Bulletin, 2017, 62(33): 61.

[21]	( 刘阳, 郭小翠, 耿金慧, 焦毅, 韩金鹏, 张志昆, 周晓, 仰大勇. 科学通报, 2017, 62(33): 61.)

[22]	Whitfield C J, Banks A M, Dura G, Love J, Fieldsend J E, Goodchild S A, Fulton D A, Howard T P. Chem. Commun., 2020, 56(52): 7108.

[23]	Ma D, Shen L, Wu K, Diehnelt C W, Green A A. Synth. Biol., 2018, 3(1): 1.

[24]	Gräwe A, Dreyer A, Vornholt T, Barteczko U, Buchholz L, Drews G, Ho U L, Jackowski M E, Kracht M, Lüders J, Bleckwehl T, Rositzka L, Ruwe M, Wittchen M, Lutter P, Müller K, Kalinowski J. PLoS One, 2019, 14(3): e0210940. DOI: 10.1371/journal.pone.0210940.

[25]	Lopreside A, Wan X Y, Michelini E, Roda A, Wang B J. Anal. Chem., 2019, 91(23): 15284.

[26]	del Valle I, Fulk E M, Kalvapalle P, Silberg J J, Masiello C A, Stadler L B. Front. Microbiol., 2021, 11: 618373. DOI: 10.3389/fmicb.2020.618373.

[27]	Amrita S, Darlin L, Akhil M, Shainlee T, Manjot P, Gulden C U. Sensors, 2018, 18(9): 2838.

[28]	Gaviria-Arroyave M I, Cano J B, Peñuela G A. Talanta Open, 2020, 2: 100006.

[29]	Voyvodic P L, Bonnet J. Curr. Opin. Biomed. Eng., 2020, 13: 9.

[30]	Zhang L Y, Guo W, Lu Y. Biotechnol. J., 2020, 15(9): 2070091.

[31]	Credou J, Berthelot T. J. Mater. Chem. B, 2014, 2(30): 4767.

[32]	Pelton R. Trac Trends Anal. Chem., 2009, 28(8): 925.

[33]	Su S X, Nutiu R, Filipe C D M, Li Y F, Pelton R. Langmuir, 2007, 23(3): 1300.

[34]	Filpponen I, Kontturi E, Nummelin S, Rosilo H, Kolehmainen E, Ikkala O, Laine J. Biomacromolecules, 2012, 13(3): 736.

[35]	Liu X J, Li X Y, Gao X, Ge L, Sun X Z, Li F. ACS Appl. Mater. Interfaces, 2019, 11(17): 15381.

[36]	Orelma H, Filpponen I, Johansson L S, Österberg M, Rojas O J, Laine J. Biointerphases, 2012, 7(1): 61.

[37]	Berlin P, Klemm D, Jung A, Liebegott H, Rieseler R, Tiller J. Cellulose, 2003, 10(4): 343.

[38]	Su S X, Ali M M, Filipe C D M, Li Y F, Pelton R. Biomacromolecules, 2008, 9(3): 935.

[39]	Pardee K, Green A A, Ferrante T, Cameron D E, DaleyKeyser A, Yin P, Collins J J. Cell, 2014, 159(4): 940.

[40]	Bracher P J, Gupta M, Mack E T, Whitesides G M. ACS Appl. Mater. Interfaces, 2009, 1(8): 1807.

[41]	Karig D K, Bessling S, Thielen P, Zhang S, Wolfe J. J. R. Soc. Interface., 2017, 14(129): 20161039.

[42]	Li X, Tian J F, Shen W. Anal. Bioanal. Chem., 2010, 396(1): 495.

[43]	Martinez A W, Phillips S T, Whitesides G M. PNAS, 2008, 105(50): 19606.

[44]	Mohidus S K G T, Wei S, Gordon W, Garnier G. Anal. Chem., 2010, 82(10): 4158.

[45]	Glavan A C, Niu J, Chen Z, Güder F, Cheng C M, Liu D, Whitesides G M. Anal. Chem., 2016, 88(1): 725.

[46]	Magro L, Jacquelin B, Escadafal C, Garneret P, Kwasiborski A, Manuguerra J C, Monti F, Sakuntabhai A, Vanhomwegen J, Lafaye P, Tabeling P. Sci. Rep., 2017, 7(1): 1.

[47]	Ali M M, Slepenkin A, Peterson E, Zhao W A. ChemBioChem, 2019, 20(7): 906.

[48]	Wang L, Chen W, Xu D, Shim B S, Kotov N A. Nano Lett., 2009, 9 (12): 4147.

[49]	Kumar S, Nehra M, Mehta J, Dilbaghi N, Marrazza G, Kaushik A. Sensors, 2019, 19(20): 4476.

[50]	Justino C, Duarte A, Rocha-Santos T. Sensors, 2017, 17(12): 2918.

[51]	Meredith N A, Quinn C, Cate D M, Reilly T H, Volckens J, Henry C S. Anal., 2016, 141(6): 1874.

[52]	Hossain S M Z, Luckham R E, McFadden M J, Brennan J D. Anal. Chem., 2009, 81(21): 9055.

[53]	Zhu C, Zhang G L, Huang Y F, Yang S M, Ren S Y, Gao Z X, Chen A L. J. Hazard. Mater., 2018, 344: 249.

[54]	Ali M M, Brown C L, Jahanshahi-Anbuhi S, Kannan B, Li Y F, Filipe C D M, Brennan J D. Sci. Rep., 2017, 7(1): 1.

[55]	Zhang Y, Zuo P, Ye B C. Biosens. Bioelectron., 2015, 68: 14.

[56]	Weng X, Neethirajan S. Microchimica Acta, 2017, 184(11): 4545.

[57]	Drummond T G, Hill M G, Barton J K. Nat. Biotechnol., 2003, 21(10): 1192.

[58]	Frew J E, Hill H A O. Anal. Chem., 1987, 59(15): 933A.

[59]	Wei H, Wu F, Yu P, Mao L Q. Chinese Journal of Analytical Chemistry, 2019, (10): 1466.

[60]	( 魏欢, 吴菲, 于萍, 毛兰群. 分析化学, 2019, (10): 1466.)

[61]	Wan K, Tao Z H, Xu L, Liu Y Q. Chinese Journal of Analytical Chemistry, 2014, 42(2): 298.

[62]	( 王昆, 陶占辉, 徐蕾, 刘亚青. 分析化学, 2014, 42(2): 298.)

[63]	Ali M M, Aguirre S D, Xu Y Q, Filipe C D M, Pelton R, Li Y F. Chem. Commun., 2009(43): 6640.

[64]	Parolo C, Merkoçi A. Chem. Soc. Rev., 2013, 42(2): 450.

[65]	Ratajczak K, Stobiecka M. Carbohydr. Polym., 2020, 229: 115463.

[66]	Takahashi M K, Xiao T, Dy A J, Dana B, Akana R T, Yoshikazu F, Nina D, Ashwin A, Collins J J. Nat. Commun., 2018, 9(1): 3347.

[67]	Suresh N, Rahin A S, Rohit C, John B, Éva N. Nanotheranostics, 2017, 1(3): 272.

[68]	Huo B Y, Hu Y L, Gao Z X, Li G K. Talanta, 2021, 222: 121565.

[69]	Xu H B, Ye Q. Journal of Food Safety & Quality, 2018, 9

[70]	( 17:113.(徐红斌, 叶青. 食品安全质量检测学报, 2018, 9(17):113.)

[71]	Kelishadi R. J. Environ. Health, 2012, 2012: 341637.

[72]	Liu Y, Wu Y H, Pang H W, Wang X X, Yu S J, Wang X K. Progress in Chemistry, 2019, (6): 831.

[73]	( 刘玥, 吴忆涵, 庞宏伟, 王祥学, 于淑君, 王祥科. 化学进展, 2019, (6): 831.)

[74]	Aljamali N, Kadhium A, Ghafil R, Jawd D S. Int. J. Prevent. Control. Ind. Pollut., 2020, 2(6): 34.

[75]	Jung J K, Alam K K, Verosloff M S, Capdevila D A, Desmau M, Clauer P R, Lee J W, Nguyen P Q, Pastén P A, Matiasek S J, Gaillard J F, Giedroc D P, Collins J J, Lucks J B. Nat. Biotechnol., 2020, 38 (12), 1451.