1. Introduction
Antimicrobial resistance (AMR) represents an escalating and pressing global menace that undermines the efficacy of antibiotics, thereby posing substantial challenges to our capacity to effectively combat bacterial infection. The excessive and inappropriate use of antibiotics hastens the emergence of drug-resistant pathogens, resulting in prolonged illnesses, elevated mortality, and escalated healthcare expenses. As traditional treatments dwindle, the imperative for innovative remedies becomes pivotal.
Responding to this exigent circumstance, both government and academia are focusing on the development of new technologies. Nanotechnology, gene technology, and molecular diagnostics have emerged as promising frontiers in the battle against antimicrobial resistance. These domains offer innovative strategies to confront the challenges presented by drug-resistant pathogens, elevating our capacity to diagnose, treat, and prevent infections. Moreover, a more encompassing perspective on healthcare innovation unlocks the potential for unconventional and collaborative approaches that hold the promise of revolutionizing antimicrobial resistance management.
In this review, our aim is to delve into these cutting-edge technologies and their potential in tackling the multifaceted issues of AMR. We will probe the role of nanotechnology in refining drug delivery precision and the potential of genetic technologies to modulate bacterial sensitivity, with the goal of uncovering the transformative possibilities inherent in these methodologies.
2. Nanotechnology In Antimicrobial Strategies
The significance of nanotechnology in antimicrobial strategies has captured significant attention due to its capacity to reshape healthcare approaches. The utilization of nanomaterials introduces a fresh perspective to addressing antimicrobial resistance (AMR) by amplifying the efficiency of diverse treatments. Nanotechnology’s relevance within healthcare hinges on its aptitude to manipulate substances at the nanoscale, enabling the creation of precise and custom-tailored intervention and shed the light on the development of novel targeted drug delivery, permitting medicines to be precisely directed into the infected regions without off-target effect. Additionally, the incorporation of nanomaterials elevates the therapeutic results of antibiotics, heightening their efficacy against tricky pathogens. This review will further discuss the applications of nanotechnology in healthcare, particularly its role in targeted drug delivery and efficacious interventions against AMR, by scrutinizing pertinent instances of successful implementation.
2.1. Application of nanotechnology in healthcare
One of the critical or significant applications of nanotechnology lies in targeted drug delivery. Nanoparticles can be engineered to encapsulate therapeutic agents and manipulate drug releasing, ensuring precise drug delivery to specific cells or tissue. In contrast to conventional methods, nano-vectors offer the advantage of concentrating a substantial number of drugs and subsequently releasing them at targeted sites to eliminate bacteria. Successful delivery of various water-soluble antibiotics, including vancomycin, gentamicin, and ampicillin, has been documented using mesoporous silica nanoparticles (MSN) [1]. Alternatively, liposomes, which resemble bacterial phospholipid bilayers, can encapsulate hydrophilic antibiotics, enabling deep penetration into bacterial cell walls for the removal of biofilm-related pathogens [2]. Addressing hydrophobic drugs, polymeric micellar structures are better suited due to their hydrophobic core functioning as a drug reservoir. A study highlighted the use of carvacrol-loaded micellar nanocarriers based on poly(oxanorborneneimide), referred to as X-NCs, achieving over 80% eradication of S. aureus and a nearly three-fold log reduction (99.5%) in its biofilm, while exhibiting no significant toxicity to co-cultured fibroblast cells [3]. To further enhance the efficiency of bacteria inhibition, a positively charged poly (ethylene glycol)-block-poly(ε-caprolactone) (PEG-b-PCL) was designed to enhance electrostatic penetration of antibiotics into acidic biofilms, preventing premature washout. Additional methods of encapsulation, including carbon nanotubes, metallic nanoparticles, HA nanoparticles, and polysaccharide nanogels, also exhibit promise in efficiently eliminating bacteria and biofilms that are drug-resistant, with each approach showcasing unique advantages and drawbacks [4].
2.2. Exploration of nanoparticles in the study of AMR
Scientists continue to actively explore the applications of nanoparticles in combating antibiotic resistance. For example, in 2023, Zhang et al. discovered a potential correlation between nanoparticle-induced bacterial memory and antibiotic resistance [5]. Nanoparticles have the capability to disrupt microbial respiration, a crucial process that provides energy for detoxification or repairing activities during periods of delay. To illustrate, the utilization of Ti3C2Tx nanosheets, a representative 2D nanomaterial from the MXene family, could incite bacterial respiration by targeting Complex I within the electron transport chai. This action creates an imbalance in NAD+ metabolism [6]. This phenomenon parallels the effect of L-serine, a compound recognized for its ability to enhance antibiotics against persisters through the elevation of intracellular NADH levels and the initiation of ROS [7]. The elevated electron transfer chain of bacterial respiration has also been observed with various other nanomaterials such as graphene, carbon nanotubes, and fullerene [7].
3. Gene Technology And Antibiotic Resistance
Gene technology has emerged as a powerful tool in the field of healthcare, offering innovative approaches to combat antibiotic resistance. In an era where the effectiveness of antibiotics is increasingly compromised by drug-resistant bacteria, gene technology provides new avenues for addressing this pressing issue. This review delves into the significance of gene technology in healthcare, particularly its role in countering antibiotic resistance. It explores how bacterial genomes can be modified through gene editing techniques to restore susceptibility to antibiotics. Furthermore, this article presents case studies that highlight the immense potential of gene technology in reshaping the landscape of antibiotic resistance and improving patient outcomes.
3.1. Introduction to gene technology and its role in healthcare
Gene technology involves the manipulation of genetic information by employing techniques such as genetic modification and gene editing. Gene technology encompasses a range of tools and methods that enable the targeted alteration of genetic information, presenting unique opportunities for innovative solutions. Gene technology has emerged as a transformative force within healthcare, with a profound impact on combating the global challenge of antimicrobial resistance (AMR). By harnessing the power of genetic manipulation, scientists can engineer microorganisms to counteract AMR. This involves introducing specific genes that encode antimicrobial peptides, enzymes, or other bioactive molecules, effectively equipping these organisms with enhanced abilities to fight drug-resistant pathogens. Beyond addressing AMR, gene technology paves the way for personalized medicine and targeted treatments. It enables the production of therapeutic proteins, enabling tailored treatments that consider individual patient characteristics. Furthermore, gene-editing techniques like CRISPR-Cas9 hold the potential to correct genetic defects and treat hereditary diseases at their source. The precision and versatility of gene technology offer a promising pathway to overcome the limitations of conventional treatments and revolutionize healthcare. As research in this field progresses, gene technology’s role in shaping a future with effective antimicrobial strategies and personalized medical interventions becomes increasingly evident, providing hope for improved patient outcomes and the mitigation of global health challenges.
3.2. Modifying bacterial genomes
The utilization of gene technology has emerged as a potent tool in the battle against antibiotic resistance. One significant avenue of this technology is the modification of bacterial genomes, which holds immense promise in addressing this global challenge.
In recent years, researchers have focused on CRISPR-Cas9. Using CRISPR-Cas9 technology, researchers can design specific guide RNA sequences, allowing the Cas9 protein to accurately cut the DNA of the target genome in a targeted manner. Once the cut occurs, the cell’s self-repair mechanism intervenes, potentially leading to changes in the genome. By controlling the repair process, researchers can achieve the addition, deletion, or modification of specific genes, thereby reversing antibiotic resistance. For instance, Wu et al. conducted sequencing and annotation of the genome from the carbapenem-resistant strain of Shewanella algae VGH117 [8]. Following this, they embarked on genome manipulation using CRISPR-Cas9 technology along with recE/recT recombinase to alter three specific candidate genes. This endeavor was undertaken with the goal of mitigating the existing carbapenem resistance within S. algae [8].
3.3. Case studies demonstrating the potential of gene technology
3.3.1. Case study 1: Antimicrobial Resistant Salmonella Concord. In recent years, the emergence of antimicrobial resistant Salmonella enterica serovar Concord (S. Concord) has been linked to severe gastrointestinal and bloodstream infections in patients from Ethiopia and Ethiopian adoptees. This serovar’s impact extends beyond Ethiopia, with sporadic cases also reported in other countries. To illuminate the evolutionary trajectory and geographical distribution of S. Concord, a comprehensive genomic analysis was conducted using 284 isolates obtained globally between 1944 and 2022 [9]. The study revealed that S. Concord is a polyphyletic serovar distributed across three distinct Salmonella super-lineages. Among these, Super-lineage A encompasses eight S. Concord lineages, four of which are associated with various countries and exhibit minimal antimicrobial resistance (AMR). However, other lineages are geographically confined to Ethiopia and display horizontally acquired resistance to a spectrum of antimicrobials commonly employed for treating invasive Salmonella infections, primarily in low- and middle-income nations. Through the assembly of complete genomes for ten representative strains, the integration of AMR markers within structurally diverse IncHI2 and IncA/C2 plasmids, as well as the chromosomal DNA, was demonstrated. The molecular surveillance of S. Concord serves as a crucial tool for comprehending AMR dynamics and facilitating a multifaceted response to the global AMR crisis. This study offers a foundational dataset for future molecular surveillance efforts [9].
3.3.2. Case study 2: Genomics-Based AMR Detection Platform. The realization of genomics’ potential to revolutionize antimicrobial resistance (AMR) identification and surveillance has posed a persistent challenge in clinical and public health microbiology. In response, an ISO-certified bioinformatics platform named abritAMR has been developed and validated for genomics-based bacterial AMR gene detection [10]. The platform harnesses the capabilities of NCBI’s AMRFinderPlus and introduces additional functionalities that categorize AMR determinants into specific antibiotic classes, generating tailored reports. Rigorous validation of abritAMR involved a comparison with PCR or reference genomes, covering an extensive range of 1500 bacterial types and 415 resistance alleles. The outcomes demonstrated the platform’s remarkable accuracy of 99.9%, sensitivity of 97.9%, and specificity of 100%. The platform’s utility was further highlighted through the comparison of genomic predictions of phenotype for 864 Salmonella spp. against agar dilution outcomes, resulting in an impressive accuracy rate of 98.9%. The implementation of abritAMR within institutional settings streamlined bioinformatics processes and reporting pathways, facilitating ease of updates and re-verification. To extend its impact, the abritAMR tool and validation datasets have been made publicly available, empowering laboratories worldwide to leverage AMR genomics effectively in their professional practices [10].
4. Molecular Diagnostics And Amr Detection
4.1. Overview of molecular diagnostics’ significance
Molecular diagnostics is critical for managing antimicrobial resistance (AMR), as it introduces a revolutionary method for the prompt identification of pathogens and genetic markers of drug resistance. This method involves using molecular biology tools to scrutinize biological markers in the genome and proteome, facilitating the identification and quantification of genetic materials and proteins linked to pathogenic organisms and drug resistance. A timely and accurate AMR diagnosis is essential for administering suitable therapeutic interventions and preventing the spread of drug-resistant pathogens. Molecular diagnostics is superior to traditional culture-based methods because it delivers higher sensitivity, specificity, and speed in producing results. This swift and precise technique is indispensable in a world where AMR is an escalating issue. The faster we can pinpoint the pathogens and their resistance patterns, the more quickly and accurately we can address the infections, thereby minimizing the morbidity and mortality associated with resistant infections. Furthermore, it helps avoid the unnecessary administration of broad-spectrum antibiotics, a major contributor to the development of resistance. Therefore, molecular diagnostics’ role in AMR management is crucial not only for individual patient care but also for public health and antibiotic stewardship at large. It also assists in the epidemiological tracking of resistant pathogens, which is vital for enacting effective infection control measures.
4.2. PCR and other molecular techniques for rapid pathogen identification
Polymerase Chain Reaction (PCR) is a commonly utilized molecular technique for the swift identification of pathogens. It amplifies particular DNA regions to create thousands to millions of copies, simplifying the identification and analysis of the target DNA sequence. PCR variants, like quantitative PCR (qPCR) and multiplex PCR, enable the quantification and concurrent detection of multiple targets. Other molecular techniques, like Next-Generation Sequencing (NGS), allow for a thorough analysis of the entire genome, facilitating the identification of novel resistance genes and mutations. After investigating the cause of diarrhea in a Cuban AIDS patient, Puebla et al. found that Cystoisospora belli is a global intestinal apicomplexan parasite that infects humans. It usually causes self-limiting infections in immunocompetent patients, but in immunocompromised individuals, it can lead to chronic, life-threatening diarrhea and dehydration. For HIV-infected patients, cystoisosporiasis is deemed an AIDS-defining illness. However, the widespread prophylactic use of TMP-SMZ against Pneumocystis jiroveci pneumonia has decreased the infection incidence in patients with accessible therapy. Typically, C. belli infection is diagnosed by identifying characteristic oocysts in stool wet mounts or modified Ziehl-Neelsen stained fecal smears. Although this parasitosis is relatively straightforward to diagnose when oocysts are excreted in high quantities, they are often excreted sporadically or in small quantities (below microscopy detection levels), as several literature-described cases have shown; in such situations, molecular approaches, especially qPCR, are beneficial [11]. Additionally, Loop-Mediated Isothermal Amplification (LAMP) offers a cost-effective, quick alternative for detecting specific DNA sequences at a stable temperature.
4.3. Detection of genetic markers of drug resistance
Detecting genetic indicators of drug resistance is essential for deciphering the resistance mechanism and directing targeted treatments. Techniques in molecular diagnostics, including PCR, NGS, and DNA microarrays, facilitate the identification of resistance genes and mutations that bestow antibiotic resistance. In 2021, Modlin and colleagues encountered a puzzling case of PZA-monoresistance in a clinical isolate. They aimed to understand the nature of this inconsistency and assess its impact on the accuracy of established and emerging targeted molecular diagnostic techniques. Targeted Sanger sequencing identified the isolate as primarily pncAWT with a minor pncASer65Ser population (both PZA-S), whereas Whole Genome Sequencing (WGS) using PacBio SMRT-sequencing identified it as a single population with a substantial deletion spanning the initial 158 nucleotides of pncA and 264 bp upstream of its start (pncAdel-264:158). After growing the sample in a PZA MGIT tube and performing targeted Sanger sequencing again, the pncA primers did not yield any sequence. Subsequently, they sequenced the drug-containing MGIT-derived sample using IonTorren, which, like SMRT-sequencing, only recovered pncAdel-264:158 [12]. For instance, alterations in the gyrA and parC genes correlate with resistance to fluoroquinolones in different bacteria, whereas the existence of the mecA gene signifies methicillin resistance in Staphylococcus aureus. Recognizing these genetic indicators allows healthcare professionals to customize the treatment plan according to the distinct resistance characteristics of the infecting pathogen, maximizing the likelihood of effective treatment.
5. Conclusion
In the urgent pursuit of countering the escalating menace of antimicrobial resistance (AMR), revolutionary technologies are poised to reshape healthcare methods and revolutionize our battle against antibiotic-resistant infections. The emergence of nanotechnology, gene technology, and molecular diagnostics represents a ray of hope, offering precise solutions to the intricate challenges posed by AMR. These innovative approaches carry the potential not only to reshape healthcare but also to significantly contribute to the worldwide efforts aimed at curbing the far-reaching consequences of AMR. However, it is pivotal to acknowledge that the way forward demands an unwavering dedication to research, collaboration, and the fostering of an environment conducive to continuous innovation.
The potency of nanotechnology in enhancing drug delivery precision and magnifying antibiotic efficacy underscores its transformative promise in addressing AMR. By capitalizing on the nanoscale domain, nanotechnology enables targeted drug administration while mitigating off-target effect to healthy tissues. Concurrently, the capacity of gene technology to manipulate microbial genomes offers a hopeful avenue for restoring susceptibility to antibiotics, presenting a glimmer of hope against resistant pathogens. Nevertheless, these strides in technology, while indispensable, constitute just a fraction of the comprehensive strategy requisite for comprehensively combating AMR. Persistent research endeavors, cooperative initiatives, and an all-encompassing approach are indispensable not only for harnessing the potential of these technologies but also for formulating a holistic framework that tackles the diverse aspects of AMR management. By embracing these innovations and cultivating a shared commitment to AMR mitigation, we usher in an era where the transformation of healthcare converges with effective AMR control, benefiting individuals and global public health alike.