Detection Of Pathogenic Bacteria Research Articles

Portable biosensors based on the CRISPR/Cas system for detection of pathogen bacteria: Up-to-date technology and future prospects

Up to now, the role of infectious pathogens bacteria in terms of severe human illnesses or even great deaths is undeniable. Over the last few years, the modern lifestyle has raised the problem of infectious pathogen bacteria more and more. Indeed, the rapid, sensitive and selective sensing platforms can prevent the spread of these microorganisms, additionally, optimize medical healthcare systems. In favor of this matter, clustered regularly interspaced short palindromic repeats (CRISPR)/Cas system, as a microbial defense system, has attracted considerable attention. To elaborate, although the system protects bacteria from being attacked by invading types, there are various types of developed biosensors using the CRISPR/Cas system for the determination of numerous targets, including bacteria. In addition, the progress of nanotechnology brings about more efficient probes and sensing approaches in terms of point-of-care (POC) detection. In favor of probes, the integration of different nanomaterials can improve stability, accuracy and sensitivity. In addition, using smartphones, microfluidics systems and lateral flow assays (LFAs) can provide portable analytical approaches for bacteria. With this regard, in this study, we aimed to comprehensively analysis current advances in portable biosensors based on the CRISPR/Cas systems and their exploitation in the quantification of pathogenic bacteria. In the first part of this study, the working principles of CRISPR/Cas systems for introducing high-potential biosensors were discussed. Afterwards, attention focused on the application of portable biosensors based on the CRISPR/Cas systems in pathogenic biosafety analysis. Eventually, the challenges and opportunities for the further progress of these types of sensing approaches were discussed.

An ultrasensitive J-shaped optical fiber LSPR aptasensor for the detection of Helicobacter pylori

The development of label-free and sensitive detection of pathogenic bacteria is of great significance for disease prevention and public health protection. In this study, an originally bent structure, named as J-shaped optical fiber probe, was first designed to engineer a localized surface plasmon resonance (LSPR) aptamer biosensor for the rapid and ultrasensitive detection of Helicobacter pylori (H. pylori). The J-shaped optical fiber probe exhibited a significant improvement in refractive index sensitivity (RIS) and LSPR signal response. Meantime, the original sequence of aptamer was truncated in order to effectively capture H. pylori on the optical fiber surface. Besides, a spacer nucleic acid with short stem-loop structure was adopted to control the aptamer density on gold nanoparticles (AuNPs) on the surface of the J-shaped optical fiber probe, which displayed a further enhancement in LSPR signal response. Benefitting from these creative designs, the proposed LSPR biosensor can realize label-free and sensitive detection of H. pylori with a detection limit as low as 45 CFU/mL and a wide linear range from 1.0 × 102 CFU/mL to 1.0 × 108 CFU/mL. At the same time, the sensing strategy can detect the pathogenic bacteria from actual water samples in one step just in 30 min without any sample pretreatment. Due to the advantages of ease-to-preparation, high sensitivity, and rapid analysis, this proposed J-shaped optical fiber LSPR aptasensor can provide a potential strategy for point-of-caring detection of pathogenic bacteria in environmental monitoring and disease diagnosis.

AIE Nanozyme-Based Long Persistent Chemiluminescence and Fluorescence for POCT of Pathogenic Bacteria.

Pathogenic bacteria are widely distributed in diverse environments and significantly threaten human health. Point-of-care testing (POCT) is a valuable way for early warnings of bacteria threat. Herein, a chemiluminescence (CL)-based ratiometric sensing platform was constructed for sensitive POCT of bacteria according to a newly designed aggregation-induced emission (AIE) molecule. The new AIE molecule presents oxidase-like properties (named as AIEzyme) and can trigger long persistent CL of luminol (LUM) with strong intensity in the absence of H2O2. The CL emission can be monitored with the naked eye for over 2 h. The emission mechanism is explored and may be attributed to the persistent reactive oxygen species generation of the AIEzyme according to the cyclic energy transfer between the AIEzyme and luminol, which catalyzes CL of luminol. Based on the CL resonance energy transfer mechanism, an afterglow luminescence system is further developed, which is used to construct a ratiometric biosensor for detection of pathogenic bacteria. With a homemade holder as a detection room and a smartphone as an analyzer, the portable biosensing platform is used for quantitative POCT of bacteria in real samples with good recovery. The detection is free of H2O2 and an external excitation source, which not only simplifies the operation but reduces interference. Specifically, the long persistent luminescence and the ratiometric strategy can significantly improve accuracy, providing an instructive way for point-of-need analysis, for example, SARS-CoV-2 detection and bioimaging analysis.

Engineering of an adaptive tandem CRISPR/Cas12a molecular amplifier permits robust analysis of Vibrio parahaemolyticus

Seeking new molecular diagnostic method for pathogenic bacteria detection is of utmost importance for ensuring food safety and protecting human health. Herein, we have engineered an adaptive tandem CRISPR/Cas12a molecular amplifier specifically designed for robust analysis of vibrio parahaemolyticus (V. parahaemolyticus), one of the most harmful pathogens. Our strategy involves the integration of three crucial processes: recombinase polymerase amplification (RPA) for copy number amplification, terminal deoxynucleotidyl transferase (TdT) for template-free strand elongation, and CRISPR/Cas12a-mediated trans-cleavage of a reporter molecule. By combining these processes, the target genomic DNA extracted from V. parahaemolyticus is able to activate many CRISPR/Cas12a units (CRISPR/Cas12an) simultaneously, resulting in a greatly amplified target signal to indicate the presence and concentration of V. parahaemolyticus. This unique model offers more advantages compared to traditional amplification models that use one RPA amplicon to activate one CRISPR/Cas12a unit. Under optimized conditions, our method enables the detection of target V. parahaemolyticus within a linear range of 1 × 102–1 × 107 CFU/mL, with an impressive limit of detection as low as 12.4 CFU/mL. It is conceivable that the adaptive tandem CRISPR/Cas12a molecular amplifier could be adapted as routine diagnostic kits in future for in-field detection of pathogens.

Rolling Circle Amplification/G‐Quadruplex‐Based Dual‐Signal Ratiometric Electrochemical Aptasensor for Ultrasensitive Detection of Pathogenic Bacteria

AbstractAccurate and efficient detection of pathogenic bacteria plays a crucial role in the diagnosis of infectious diseases. However, rapid and highly sensitive detection of specific bacteria in clinical samples remains challenging. In this study, we developed a ratiometric dual‐signal electrochemical biosensor for ultrasensitive detection of pathogenic bacteria, based on an aptamer recognition‐induced rolling circle amplification (RCA)/G‐quadruplex strategy. On the surface of a gold electrode chip, we used an aptamer sequence (P1) conjugated to ferrocene (Fc) to capture the target bacterium. This capture released a different, previously bound sequence (P2) sequence that initiated the RCA/G‐quadruplex cascade, which, in the presence of potassium ions, was able to bind the electrochemical indicator methylene blue (MB). We integrated the signals from the loss of Fc and the appearance of MB (IMB/IFC ratio) to quantify the target bacteria concentration. This biosensor showed excellent detection performance and specificity, with a detection limit of 10 CFU/mL. Notably, it showed great diagnostic potential for clinical infectious diseases caused by pathogenic bacteria in precise/personalized medicine applications.

Detection of Escherichia coli by capillary electrophoresis assisted by large volume sample stacking and nicking endonuclease signal amplification

Efficient, accurate and economical detection of pathogenic bacteria is crucial in ensuring food safety and preventing foodborne illnesses. In this study, a capillary electrophoresis coupled laser-induced fluorescence assay (CE-LIF) was developed for the detection of Escherichia coli (E. coli) by detecting its specific DNA. The CE-LIF was assisted by both online enrichment and offline amplification to improve the detection sensitivity of bacterial DNA. Here the online amplification was performed by large volume sample stacking (LVSS), while the offline amplification was nicking endonuclease signal amplification (NESA). Under the optimal experimental conditions, the detection limit of bacterial target DNA was 2.5 fM, and the conversion concentration of E. coli was 3 CFU · mL−1. The method had been applied to the detection of commercially available skim milk samples with good results, which proved that it could be used as an effective tool for food and environmental bacteria monitoring.

Urchin-Shaped Au-Ag@Pt Sensor Integrated Lateral Flow Immunoassay for Multimodal Detection and Specific Discrimination of Clinical Multiple Bacterial Infections.

A new lateral flow immunoassay strip (LFIA) combining sensitive detection and identification of multiple bacteria remains a huge challenge. In this study, we first developed multifunctional urchin-shaped Au-Ag@Pt nanoparticles (UAA@P NPs) with a unique combination of colorimetric-SERS-photothermal-catalytic (CM/SERS/PT/CL) properties and integrated them with LFIA for multiplexed detection and specific discrimination of pathogenic bacteria in blood samples. Unlike the conventional LFIA that relied on antibody (Ab), this novel LFIA introduced 4-mercaptophenylboronic acid (4-MPBA) as an ideal Ab replacer that was functionalized on UAA@P NPs (UAA@P/M NPs) with outstanding binding and enrichment capacities toward bacteria. Taking Staphylococcus aureus (S. aureus) as model bacteria, the limit of detection (LOD) was 3 CFU/mL for SERS-LFIA, 27 CFU/mL for PT-LFIA, and 18 CFU/mL for CL-LFIA, three of which were over 330-fold, 37-fold, and 55-fold more sensitive than ordinary visual CM-LFIA, respectively. Besides, this SERS-LFIA is capable of identifying three types of bacterial spiked blood samples (E. coli, S. aureus, and P. aeruginosa) effectively according to specific bacterial Raman "fingerprints" by partial least-squares-discriminant analysis (PLS-DA). More importantly, this LFIA was successfully applied to blood samples with satisfactory recoveries from 90.3% to 108.8% and capable of identifying the infected patients (N = 4) from healthy subjects (N = 2) with great accuracy. Overall, the multimodal LFIA incorporates bacteria discrimination and quantitative detection, offering an avenue for early warning and diagnosis of bacterial infection.

Magnetic relaxation switching biosensor for one-step detection of Vibrio parahaemolyticus based on click chemistry-mediated sol-gel system

In this study, a magnetic relaxation switching (MRS) biosensor was developed for one-step detection of pathogenic bacteria Vibrio parahaemolyticus (VP). The biosensor is based on a click chemistry-mediated sol-gel system and utilizes low-field nuclear magnetic resonance (LF NMR) detection. VP aptamer (Apt)-coated hollow mesoporous silica microspheres (HMSMs) with saturated sodium ascorbate (SAA) solution sealed inside, HMSMs@SAA&Apt, are designed as the signal unit. When the target VP is present, the Apt binds to VP and releases SAA from the signal unit. The released SAA reduces Cu2+ in the test solution to Cu+, which catalyzes the click chemistry reaction between azide-modified polyethylene glycol (PEG-Azide) and acetylene-modified polyacrylic acid (PAA-Alkyne). The sol-gel transition is triggered, a large amount of "free" water is converted into "bound" water, resulting in the decrease of transverse relaxation time (T2). Under the optimal experimental conditions, the variation of T2 (ΔT2) was linearly related to the logarithm of VP concentration in the range of 10 ∼ 1.0 × 108 CFU/mL, with a limit of detection of 5 CFU/mL. The assay demonstrated good selectivity, stability, and reproducibility. It is completed in one step, holding promising applications in the rapid field detection of pathogenic bacteria.

Lateral Flow Strip Biosensors for Foodborne Pathogenic Bacteria via Direct and Indirect Sensing Strategies: A Review.

Rapid and sensitive detection of foodborne pathogenic bacteria is particularly important for the prevention and control of foodborne diseases. The lateral flow strip biosensor (LFSB) is one of the most promising point-of-care detection tools and has been widely used in food safety monitoring. This review introduces recent advances in the detection of foodborne pathogenic bacteria using LFSBs. According to different bacterial biomarkers, we summarize the direct and indirect sensing strategies of bacterial LFSBs. The direct sensing strategies for whole bacterial cells are divided into antibodies, antibody alternatives, and label-free according to the recognition elements. The indirect sensing strategies refer to the detection of bacterial nucleic acids and metabolites. Next, we compare and discuss the applications of direct and indirect sensing strategies. Finally, the existing challenges, future perspectives, and development directions are discussed, which will facilitate the theoretical innovation and practical application for bacterial LFSBs.

Portable biosensor-based oral pathogenic bacteria detection for community and family applications.

Detection of oral pathogens is essential in the management of oral diseases, as their occurrence and progression are closely linked to an imbalance in these microorganisms. Detection techniques such as microbial cultures, enzyme-linked immunosorbent assays and polymerase chain reactions are highly dependent on complex testing procedures and specialized laboratory equipment, making prevention and early diagnosis of oral diseases difficult. To comprehensively implement oral disease prevention and early diagnosis in social groups, there is an urgent need for portable testing methods for oral pathogenic bacteria that can be applied in community and home settings. In this review, several common portable biosensors for pathogenic bacteria are first described. Based on the goal of achieving primary prevention and diagnosis of oral diseases, we elaborate and summarize portable biosensors for common oral pathogenic bacteria in terms of how to achieve portability of the technique. This review aims to reflect the current status of portable biosensors for common oral pathogens and to lay the foundation for the further realization of portable detection of oral pathogens.

Microbiological Detection of Pathogenic Bacteria in Diabetic Foot Ulcers of Type 1 and 2 Diabetic Mellitus Patients

Globally, diabetes mellitus (DM) is a serious health issue. Diabetic neuropathy, the most prevalent micro-vascular consequence of diabetes, raises the chance of developing foot ulcers. Different aerobic bacteria, including Gram positive cocci, Staphylococcus aureus, and (beta-haemolytic) Streptococcus are some of the pathogens that can colonize DFUs. Coagulation-negative Staphylococcus (CNS), Corynebacterium, Enterococcus, and Cutibacteria are frequently found in ulcer cultures in clinical practice; however they are typically regarded as colonizing rather than harmful bacteria. Anaerobes are detected in a smaller minority of DFIs and have been associated to altered innate and adaptive immune responses in the hyperglycemic milieu. Gram-negative bacteria account for about one third of DFIs. About 70 participants in all, including 29 individuals with type 1 diabetes and 41 patients with type 2 diabetes who have diabetic foot ulcers. Scarping of ulcers takes place after regular saline wound cleansing to eliminate surface pollutants, and then exudate is sampled. Pus, or discharges from the base of the ulcer, are then submitted to the laboratory as quickly as possible using an aseptic approach. Then, the bacteria were inoculated onto various culture media, such as MacConkey and Blood agar as enrichment media, and incubated aerobically at 37°C for 24 hours to isolate aerobic bacteria. The following day, the bacteria were checked for growth, and a pure colony was then prepared. According to morphological, microscopic, and particular cards of the automated VITEK2 system, isolated bacteria were identified.

A CRISPR/dCas9‐enabled, on‐site, visual, and bimodal biosensing strategy for ultrasensitive and self‐validating detection of foodborne pathogenic bacteria

AbstractIt is imperative to develop practicable pathogenic bacteria detection methods. We devised a biosensor for the ultrasensitive detection of Salmonella typhimurium (S. typhi), termed as SCOUT‐dCas9 (ultrasensitive, cross‐validating, on‐site, and dUal‐mode test using CRISPR/dCas9). Simply, the species‐specific invA gene of S. typhi was amplified using loop‐mediated isothermal amplification with a biotinylated primer, which can be specifically “pulled down” by “antibody‐like” dCas9‐single guide RNA to form a ternary complex. SYBR Green I and streptavidin‐modified alkaline phosphatase were used to functionalize them to generate fluorescent and colorimetric signals, respectively. With this strategy, the target could be dexterously converted into bimodal signals that were cross‐validated to afford more reliable results. For both modes, SCOUT‐dCas9 was able to detect as low as 1 CFU/mL with a dynamic range from 1 to 109 CFU/mL. Lastly, SCOUT‐dCas9 had satisfactory selectivity and was capable of detecting S. typhi‐contaminated real food samples. SCOUT‐dCas9 provides a robust platform for bacterial detection.

A visual colorimetric assay based on phage T156 and gold nanoparticles for the sensitive detection of Salmonella in lettuce

In this study, a new technique was developed for visual and precise identification of Salmonella using phage T156-mediated aggregation of gold nanoparticles. The phage binds to gold nanoparticles in a dispersed and stable state under high NaCl concentrations. When Salmonella is introduced, the phage specifically recognizes and adsorbs the targeted bacteria, causing the AuNPs to undergo a discoloration reaction resulting in aggregation, which enables Salmonella visualization. The method has a detection range of 3.8 × 101–3.8 × 109 CFU/mL and a limit of detection of 38 CFU/mL and can produce results in approximately 80 min. The technique was also tested on field samples, including spiked lettuce, and was found to be accurate with a recovery rate of 81.0–119.2% and relative standard deviations ranging from 3.3% to 14.7%. Notably, this technique utilizes phages as recognition elements in colorimetric methods, offering simplicity, speed, and the ability to effectively distinguish between live and dead Salmonella. It demonstrates remarkable sensitivity, specificity, and accuracy. Furthermore, it presents a novel avenue for the rapid detection of other pathogenic bacteria.

Ag+-gated peroxidase activity of gold nanoparticles for sensitive detection of Escherichia coli

Escherichia coli (E. coli) is one of the most ubiquitous foodborne pathogens that can cause infections and threaten human health. Herein, a colorimetric method for sensitive detection of E. coli was established by using enzyme-nanozyme cascade reaction for signal amplification. Gold nanoparticles (AuNPs) are well-known nanozymes due to their high peroxidase-like activity. When the dense cetyltrimethylammonium bromide (CTAB) membrane on the surfaces of AuNPs kept the substrate away from AuNPs, the peroxidase activity of AuNPs was inhibited. However, the CTAB membrane could be disrupted by Ag+, resulting in enhanced peroxidase activity of AuNPs. When E. coli was present, the enzyme-nanozyme cascade reaction was initiated. The substrate p-aminophenyl β-D-galactopyranoside (PAPG) was hydrolyzed to the reductive p-aminophenol (PAP) by beta-galactosidase (β-gal) in E. coli, reducing Ag+ to Ag. Consequently, CTAB-AuNPs remained weak peroxidase activity and could not catalyze the H2O2-mediated oxidation of TMB. As the amount of E. coli increased, the absorbance of TMB decreased along with a color change from deep blue to pink. The absorbance intensity displayed a linear dependence on E. coli from 1.0 × 102 to 1.0 × 109 CFU mL−1. Therefore, the proposed method holds good prospects in foodborne pathogenic bacteria detection.

Dual targets-induced specific hemin/G-quadruplex assemblies for label-free electrochemical detection capable of distinguishing Salmonella and its common serotype in food samples

Efficient detection of pathogenic bacteria is paramount for ensuring food safety and safeguarding public health. Herein, we developed a label-free and signal-on dual-target recognition electrochemical DNA sensing platform based on the conformational formation of split G-quadruplex. This platform focused on achieving sensitive and low-cost detection of Salmonella and its most human-infecting S. typhimurium serotype. In simple terms, the dual-target recognition probe (DTR-6P) was ingeniously designed for the loop sequence on the loop-mediated isothermal amplification (LAMP) amplicons. It could recognize two different genes and release their corresponding G-rich sequences. The exfoliated G-rich sequences could be captured by the capture probes on the electrode, and then the bimolecular G-quadruplex or the tetramolecular G-quadruplex would be formed to capture hemin, thereby enabling dual-signal reporting. The minimum detection amount of target genes can be as low as 2 copies/μL. Encouragingly, the real food samples contaminated by Salmonella and the S. typhimurium serotype can be readily identified. The sensing platform with ingenious design paves a new way for label-free, multi-target simultaneous detection, whose advantage of rapidity, sensitivity, cost-effectiveness, and specificity also lay a solid foundation for practical applications.

Target enrichment of uncultured human oral bacteria with phage-derived molecules found by single-cell genomics

Advances in culture-independent microbial analysis, such as metagenomics and single-cell genomics, have significantly increased our understanding of microbial lineages. While these methods have uncovered a large number of novel microbial taxa, many remain uncultured, and their function and mode of existence in the environment are still unknown. This study aims to explore the use of bacteriophage-derived molecules as probes for detecting and isolating uncultured bacteria. Here, we proposed multiplex single-cell sequencing to obtain massive uncultured oral bacterial genomes and searched prophage sequences from over 450 obtained human oral bacterial single-amplified genomes (SAGs). The focus was on the cell wall binding domain (CBD) in phage endolysin, and fluorescent protein-fused CBDs were generated based on several CBD gene sequences predicted from Streptococcus SAGs. The ability of the Streptococcus prophage-derived CBDs to detect and enrich specific Streptococcus species from human saliva while maintaining cell viability was confirmed by magnetic separation and flow cytometry. The approach to phage-derived molecule generation based on uncultured bacterial SAG is expected to improve the process of designing molecules that selectively capture or detect specific bacteria, notably from uncultured gram-positive bacteria, and will have applications in isolation and in situ detection of beneficial or pathogenic bacteria.

A dual-mode ratiometric aptasensor for accurate detection of pathogenic bacteria based on recycling of DNAzyme activation

Pathogenic bacteria have a significant impact on food safety. Herein, an innovative dual-mode ratiometric aptasensor was constructed for ultrasensitive and accurate detection of Staphylococcus aureus (S. aureus) based on recycling of DNAzyme activation on gold nanoparticles-functionalized MXene nanomaterials (MXene@Au NPs). Electrochemiluminescent (ECL) emitter-labeled probe DNA (probe 2-Ru) containing the blocked DNAzyme was partly hybridized with aptamer and then captured by electrochemical (EC) indicator-labeled probe DNA (probe 1-MB) on electrode surface. When S. aureus presented, the conformation vibration of probe 2-Ru activated the blocked DNAzymes, leading to recycling cleavage of probe 1-MB and ECL tag close to electrode surface. Based on the reverse change tendencies of ECL and EC signals, aptasensor achieved S. aureus quantification from 5 to 108 CFU/mL. Moreover, the self-calibration characteristic of the aptasensor with dual-mode ratiometric readout ensured the reliable measurement of S. aureus in real samples. This work showed useful insight into sensing foodborne pathogenic bacteria.

NIR II light response-based PDA/AuPt@CuS composites: Simultaneous readout of temperature and pressure sensing strategy for portable detection of pathogenic bacteria

In this study, we developed a simultaneous readout of pressure and temperature dual-signals platform based on the second near-infrared (NIR II) light response-based polydopamine (PDA)-functionalized-AuPt nanoparticles (NPs)@CuS nanosheets (PDA/AuPt@CuS NS) composite. Due to the excellent NIR photothermal performance of PDA/AuPt@CuS NS, it contribute to the decomposition of H2O2 and NH4HCO3 to generate gases (including O2, CO2, and NH3) can be promoted, which can amplify the pressure signals in a sealed container. A sandwich mode is formed between Fe3O4 NPs and PDA/AuPt@CuS NS based on the dual-aptamer when target pathogenic bacteria is present. And, it is possible to convert the molecular recognition signals between the dual-aptamers into amplified pressures and temperatures, which can be read out by a portable pressure meter and smartphones simultaneously. It may offer the possibility for quantitative POCT analysis of Pathogenic Bacteria. Moreover, because of the high photothermal efficiency of this method, the developed dual-mode method can achieve that following the detection of bacteria and killing them immediately. As a result, secondary contamination is eliminated and bacterial transmission is avoided. The developed dual-signal sensing platform is also inexpensive, simple to operate and rapidly, indicating that it can be used for food safety analysis, clinical applications, and environmental monitoring.

Polymeric and Paper-Based Lab-on-a-Chip Devices in Food Safety: A Review.

Food quality and safety are important to protect consumers from foodborne illnesses. Currently, laboratory scale analysis, which takes several days to complete, is the main way to ensure the absence of pathogenic microorganisms in a wide range of food products. However, new methods such as PCR, ELISA, or even accelerated plate culture tests have been proposed for the rapid detection of pathogens. Lab-on-chip (LOC) devices and microfluidics are miniaturized devices that can enable faster, easier, and at the point of interest analysis. Nowadays, methods such as PCR are often coupled with microfluidics, providing new LOC devices that can replace or complement the standard methods by offering highly sensitive, fast, and on-site analysis. This review's objective is to present an overview of recent advances in LOCs used for the identification of the most prevalent foodborne and waterborne pathogens that put consumer health at risk. In particular, the paper is organized as follows: first, we discuss the main fabrication methods of microfluidics as well as the most popular materials used, and then we present recent literature examples for LOCs used for the detection of pathogenic bacteria found in water and other food samples. In the final section, we summarize our findings and also provide our point of view on the challenges and opportunities in the field.

Poly(amino acids) towards sensing pathogenic bacteria: A review

On the planet Earth, the living of numerous species has strongly depended on microorganisms, one of which is bacteria. Among various bacteria, the role of pathogenic bacteria in terms of prevalent diseases, environmental disasters, and even animal life are undeniable. Indeed, in the modern era, diagnosis of the spread of these bacteria in different settings including the human body, food matrix, and environment can able us to control them. Despite the advancement of sensing platforms, there is a huge shortage in the development of sensitive and selective sensing approaches. Hence, fabrication efficient sensing-assay can play an important role. Amino acids (AAs) are part of biocompatible materials that have attracted attention as a promising class in various biosensors as sensing probes. This review focuses on the application of poly(AAs) such as cysteine and poly (Cysteine), lysine and poly (Lysine), glutamic acid and poly (Glutamic acid), glycine and poly (Glycine) and other types of poly(AAs) in terms of pathogenic bacteria detection. In addition, the role of different nanomaterials along with biological factors in increasing the sensitivity and selectivity of these types of assays will be reviewed. To elaborate, the identification of pathogenic bacteria will be investigated using different diagnostic platforms such as electrochemiluminescence (ECL), optical, electrochemical (EC), and photoelectrochemical (PEC) techniques, which will pay special attention to the role of AA and poly AA. Finally, the existing technological and research gaps will be examined and criticized in order to solve them in the future. It should be noted the prepared study is the first comprehensive review of bacterial assay detection by using poly(AAs) in the construction of biosensors.