Rapid Diagnosis Of Bacterial Infections Research Articles

Aeromonas veronii specific aptamer and peroxidase mimic tyrosine-capped gold NanoZymes enable highly specific sensing of fish pathogenic bacteria

Despite major advances in biosensing, quick, dependable, and effective on-site detection of bacterial infections remains a serious issue, owing to a lack of acceptable or appropriate diagnostic platforms. To address this gap, we presented a new colorimetric gold NanoZyme aptasensor for rapid sensing of Aeromonas veronii, an infectious bacterial disease in fish. The A. veronii-specific aptamer (AVS01) was developed through Cell-SELEX. The sensing mechanism involves inhibition of AuNPs induced peroxidase-mimic catalytic activity through surface adsorption by AVS01 which in the presence of the A. veronii desorb from the AuNPs allowing recovery of the catalytic activity leading to colorimetric response, whereas the sensor is insesnsitive to other nontarget bacterial cells. This method is very specific and sensitive, allowing for the quick and visible sensing of A. veronii with a detection limit of 1281 CFU mL−1 within 15 min. The method has great potential for rapid diagnosis of bacterial infection in fish caused by A. veronii.

Electrochemical biosensors for rapid diagnosis of bacterial infections: Design, targets and applications in clinical setting

AbstractIn clinical settings, quickly identifying and detecting bacterial infections is crucial. Current methods for identifying and measuring these diseases take a long time and are frequently expensive. Hence, biosensor systems may be a viable alternative to conventional systems. Electrochemical systems based on different materials have great potential for developing rapid methods to detect pathogens in many areas. They are cheaper and reliable, have a relatively fast response time, and provide better stability and sensitivity. This review article assesses the recent advancements and potential of electrochemical biosensors for monitoring/detecting different pathogens in clinical samples. After mentioning standard techniques for detecting various microorganisms, the article introduced the biosensor‘s fundamental conceptualization and its varieties, functioning principles, and advantages. Then it discussed the applicability of various electrochemical systems employed in clinical analysis. It reviewed DNA, RNA and protein‐based systems as methods of direct pathogen analysis and underlined metabolite‐based indirect systems, the new trend in microbiology and life sciences, with their potential for detecting infectious diseases.

Nötrofil Hacim, İletkenlik ve Işık Saçılımı Parametrelerinin Bakteriyel Enfeksiyonlarda Hızlı Tanı Aracı Olarak Kullanımının Araştırılması

Aim: Early diagnosis of bacterial infections is crucial in planning treatment. Thus, it is important to determine the gram staining property of bacteria as well as the presence of bacterial infection. The acute bacterial infection leads to morphological changes in neutrophils. In this study, we investigated the use of neutrophil volume, conductivity and light-scattering (VCS) parameters as potential indicators for rapid diagnosis of bacterial infections, and to distinguish infections caused by gram-positive and gram-negative bacteria.
 Material and Methods: Patients with urinary tract infections, pneumonia, wound site infections and sepsis were included. The control group comprised patients without bacterial infections. The blood samples of the patients were examined for white blood cell and neutrophil counts; neutrophil VCS parameters were determined using a Coulter Analyzer. The VITEK 2 Compact System was used to detect microbial growth.
 Results: The blood sample data of 472 patients sent to our laboratory for Complete Blood Count analyses from various clinics were analyzed in this study. A total of 370 samples showed significant growth in their bacteriological culturing, whereas the remaining 102 samples showed no growth. For the detection of bacterial growth, the specificities of median neutrophil volume (MNV) and median neutrophil conductivity (MNC) were found to be 96% and 99%, respectively. In addition, median neutrophil light scattering (MNS) was higher in the gram-negative bacterial group than in the gram-positive bacterial group. 
 Conclusion: The use of neutrophil VCS parameters is an effective and time-saving method to identify bacterial infections and distinguish between gram-positive and gram-negative bacterial infections.

Early appropriate diagnostics and treatment of MDR Gram-negative infections.

The term difficult-to-treat resistance has been recently coined to identify Gram-negative bacteria exhibiting resistance to all fluoroquinolones and all β-lactam categories, including carbapenems. Such bacteria are posing serious challenges to clinicians trying to identify the best therapeutic option for any given patient. Delayed appropriate therapy has been associated with worse outcomes including increase in length of stay, increase in total in-hospital costs and ∼20% increase in the risk of in-hospital mortality. In addition, time to appropriate antibiotic therapy has been shown to be an independent predictor of 30 day mortality in patients with resistant organisms. Improving and anticipating aetiological diagnosis through optimizing not only the identification of phenotypic resistance to antibiotic classes/agents, but also the identification of specific resistance mechanisms, would have a major impact on reducing the frequency and duration of inappropriate early antibiotic therapy. In light of these considerations, the present paper reviews the increasing need for rapid diagnosis of bacterial infections and efficient laboratory workflows to confirm diagnoses and facilitate prompt de-escalation to targeted therapy, in line with antimicrobial stewardship principles. Rapid diagnostic tests currently available and future perspectives for their use are discussed. Early appropriate diagnostics and treatment of MDR Gram-negative infections require a multidisciplinary approach that includes multiple different diagnostic methods and further consensus of algorithms, protocols and guidelines to select the optimal antibiotic therapy.

Utility of Procalcitonin Over C-Reactive Protein and WBC as an Early and Rapid Diagnostic Marker of Bacterial Infections in Febrile Patients with or no Hemoculture Results as Gold Standard in a Malaria Endemic Zone of Cameroon

Background & objectives: Early identification of bacterial infection in patients with fever is important for prompt and specific treatment. However, the available biomarkers such as C-reactive protein (CRP) and leukocyte counts are not specific to bacterial diagnosis. This study aimed to assess the diagnostic value of procalcitonin (PCT) over CRP and leukocyte counts for bacterial infection screening in febrile patients while awaiting their hemoculture results, which takes up to 7 Days before results are available though it is the gold standard for the diagnosis of bacterial infections. Methods: Blood samples were collected from febrile patients between January and July 2020 then processed for blood cultures. PCT, CRP and WBC levels were measured. The patients were divided into two groups according to the final diagnosis: bacterial infection group (group1) and non-bacterial infection group (group 2).SPSS version 20.0 software package was used for data analysis and normally distributed variables were calculated using mean, standard deviation and ANOVA test while median and range were used for variables without a normal distribution. Significance testing was done using Kruskal-Wallis h test and Wilcoxon two sample test and the diagnostic accuracy was assessed by calculating the area (AUC) under the receiver operating characteristic curve (ROC). Results: There were significant (P<0.05) difference in the levels of PCT, CRP and WBC among the two groups. The PCT levels of patients in the bacterial infections group were significantly higher than those in the nonbacterial infections group (27.9 vs., 11.7 P < 0.001). The best cut-off value to detect bacterial infections was 1.46 ng/ml for PCT. PCT, CRP and WBC had areas under the curve of 0.71, 0.66 and 0.45 respectively and sensitivity of 100%, 72.7% and 27.3% respectively. Interpretation & conclusions: Our results showed that PCT was a valuable marker for the early and rapid diagnosis of bacterial infections in febrile patients in our setting when compared to CRP and WBC. However, prospective and large scale studies are warranted to confirm these findings in Cameroon.

New Microbiological Techniques for the Diagnosis of Bacterial Infections and Sepsis in ICU Including Point of Care.

Purpose of ReviewThe aim of this article is to review current and emerging microbiological techniques that support the rapid diagnosis of bacterial infections in critically ill patients, including their performance, strengths and pitfalls, as well as available data evaluating their clinical impact.Recent FindingsBacterial infections and sepsis are responsible for significant morbidity and mortality in patients admitted to the intensive care unit and their management is further complicated by the increase in the global burden of antimicrobial resistance. In this setting, new diagnostic methods able to overcome the limits of traditional microbiology in terms of turn-around time and accuracy are highly warranted. We discuss the following broad themes: optimisation of existing culture-based methodologies, rapid antigen detection, nucleic acid detection (including multiplex PCR assays and microarrays), sepsis biomarkers, novel methods of pathogen detection (e.g. T2 magnetic resonance) and susceptibility testing (e.g. morphokinetic cellular analysis) and the application of direct metagenomics on clinical samples. The assessment of the host response through new “omics” technologies might also aid in early diagnosis of infections, as well as define non-infectious inflammatory states.SummaryDespite being a promising field, there is still scarce evidence about the real-life impact of these assays on patient management. A common finding of available studies is that the performance of rapid diagnostic strategies highly depends on whether they are integrated within active antimicrobial stewardship programs. Assessing the impact of these emerging diagnostic methods through patient-centred clinical outcomes is a complex challenge for which large and well-designed studies are awaited.

AB026. Gene expression for disease stratification and prognostication in neonatal encephalopathy

: Neonatal encephalopathy is a leading cause of neonatal death and lifelong disability accounting for 1 million deaths every year with 99% of the disease burden in developing countries. Most putative neuroprotectants, including cooling therapy, are effective only when initiated within few hours of birth, hence early identification of ‘at risk’ encephalopathic infants is vital. Magnetic resonance imaging and spectroscopy are considered gold standard for predicting long-term adverse outcomes after neonatal encephalopathy. However, they are limited by the inability to perform a scan as soon as possible after birth. Inevitably, clinicians have to make treatment decisions before this information is available. Therefore, it is vitally important to identify babies at risk of long-term neurodisability as early as possible after birth, to initiate preventative therapy. In the past decade, genomic and transcriptomic signatures have revolutionized personalised chemotherapy, particularly in breast cancer. Our group and others have effectively exploited host transcriptomic profiling for the rapid diagnosis of bacterial infection and Kawasaki disease, and for disease stratification in tuberculosis. We have recently shown that babies with neonatal encephalopathy have a different gene expression profile, when compared with age matched healthy newborn babies and that transcriptomic signatures at birth can identify the babies who develop adverse outcomes with high accuracy. These findings highlight that gene expression hold great potential as prognostic and treatment decision biomarker.

1758. Impact of Accelerate Pheno™ Rapid Blood Culture Detection System on Laboratory and Clinical Outcomes in Bacteremic Patients

BackgroundMolecular-based automated systems for the rapid diagnosis of bacterial infections have potential to improve patient care. The Accelerate Pheno™ blood culture detection system (ACCEL) is an FDA approved platform that allows for identification (ID) and antimicrobial susceptibility testing (AST) 8 hours following growth in routine culture.MethodsThis is a single-center retrospective chart review of bacteremic adult inpatients before and after implementation of ACCEL. Laboratory and clinical data were collected February–March 2018 (intervention) and compared with a January–April 2017 historical cohort (standard of care). Standard of care ID and AST were performed using VITEK® MS (MALDI-TOF MS) and VITEK®2, respectively. An active antimicrobial stewardship program was in place during both study periods. Patients with polymicrobial cultures, off-panel isolates, previous positive culture, or who were discharged prior to final AST report were excluded. Primary outcome was length of stay (LOS). Secondary outcomes were inpatient antibiotic duration of therapy (DOT) and time to optimal therapy (TTOT). Nonparametric unadjusted analyses were performed due to non-normal distributions. Statistics were performed using SAS 9.4.ResultsOf the 143 positive cultures performed on ACCEL during intervention, 118 (83%) were identified as on-panel organisms. Seventy-five (64%) of these 118 cultures and 79 (70%) of 113 reviewed standard of care cultures met inclusion criteria. Patient comorbidities (P = NS), MEWS severity score (P = 0.10), source of bacteremia (P = NS), and pathogen detected (P = 0.30) were similar between cohorts. Time from collection to ID (28.2 ± 12.7 hours vs. 53.8 ± 20.9 hours; P < 0.001) and AST (31.9 ± 11 hours vs. 71.8 ± 20 hours; P < 0.001) were shorter in the intervention arm.Clinical OutcomesStandard of Care (Mean ± SD) N = 79Intervention (Mean ± SD), N = 75 P-valueLOS (days)12.1 (11.9)9.1 (7.6)0.03TTOT (hours)73.5 (50.2)37.5 (32.7)<0.001Total antibiotic DOT (days)9.0 (7.5)7.0 (4.6)0.05Meropenem DOT (days)6.6 (3.7)3.7 (2.1)0.03ConclusionCompared with standard of care, ACCEL shortens laboratory turn-around-time and improves clinical outcomes. The use of this system has resulted in decreased mean antibiotic DOT, TTOT, and LOS. Further studies are needed to verify these findings.Disclosures All authors: No reported disclosures.

Surface-Enhanced Raman Scattering for Rapid Detection and Characterization of Antibiotic-Resistant Bacteria.

As the prevalence of antibiotic-resistant bacteria continues to rise, biosensing technologies are needed to enable rapid diagnosis of bacterial infections. Furthermore, understanding the unique biochemistry of resistance mechanisms can facilitate the development of next generation therapeutics. Surface-enhanced Raman scattering (SERS) offers a potential solution to real-time diagnostic technologies, as well as a route to fundamental, mechanistic studies. In the current review, SERS-based approaches to the detection and characterization of antibiotic-resistant bacteria are covered. The commonly used nanomaterials (nanoparticles and nanostructured surfaces) and surface modifications (antibodies, aptamers, reporters, etc.) for SERS bacterial detection and differentiation are discussed first, and followed by a review of SERS-based detection of antibiotic-resistant bacteria from environmental/food processing and clinical sources. Antibiotic susceptibility testing and minimum inhibitory concentration testing with SERS are then summarized. Finally, recent developments of SERS-based chemical imaging/mapping of bacteria are reviewed.

Multiplex quantification of Escherichia coli, Salmonella typhi and Vibrio cholera with three DNA targets in single reaction assay

Escherichia coli (E. coli), Salmonella typhi and Vibrio cholera harmful pathogens, which causes various diseases in humans. Rapid diagnosis of bacterial infection is an important for patient management and appropriate therapy during the early phase of the bacterial infected diseases. Among the existing techniques for identifying pathogens were less sensitive and time-consuming processes. In the present study total, 48 clinical 31 blood and 17 urine samples of patients suspected with the infections were collected from SVRR Hospital and used to detect the pathogens. Multiplex polymerase chain reaction (PCR) assay was set to design for the identification of Escherichia coli, Salmonella typhi and Vibrio cholera from the different clinical samples. Rapid diagnosis of Escherichia coli (E. coli), Salmonella and Vibrio cholera pathogens can be done with simultaneously in a single multiplex PCR assay by using specific primers with adjusted PCR conditions. Through this approach, the results represented with out of 31 blood samples 1–15 shows the positive with E. coli and remaining 14 only 11 were correlated with multiplex results of Vibrio cholera, remaining the urine samples all are positive with 17 samples correlate with the Salmonella typhi. Through the high specificity benefits of excellent sensitivity, with high resolution and reproducibility. This method of results proved and illustrates the best potential system for diagnosing the infectious disease with modern trendy.

Streaming algorithms for identification of pathogens and antibiotic resistance potential from real-time MinION(TM) sequencing.

The recently introduced Oxford Nanopore MinION platform generates DNA sequence data in real-time. This has great potential to shorten the sample-to-results time and is likely to have benefits such as rapid diagnosis of bacterial infection and identification of drug resistance. However, there are few tools available for streaming analysis of real-time sequencing data. Here, we present a framework for streaming analysis of MinION real-time sequence data, together with probabilistic streaming algorithms for species typing, strain typing and antibiotic resistance profile identification. Using four culture isolate samples, as well as a mixed-species sample, we demonstrate that bacterial species and strain information can be obtained within 30 min of sequencing and using about 500 reads, initial drug-resistance profiles within two hours, and complete resistance profiles within 10 h. While strain identification with multi-locus sequence typing required more than 15x coverage to generate confident assignments, our novel gene-presence typing could detect the presence of a known strain with 0.5x coverage. We also show that our pipeline can process over 100 times more data than the current throughput of the MinION on a desktop computer.Electronic supplementary materialThe online version of this article (doi:10.1186/s13742-016-0137-2) contains supplementary material, which is available to authorized users.

Label-free bimodal waveguide immunosensor for rapid diagnosis of bacterial infections in cirrhotic patients

Spontaneous bacterial peritonitis is an acute bacterial infection of ascitic fluid; it has a high incidence in cirrhotic patients and it is associated with high mortality. In such a situation, early diagnosis and treatment is crucial for the survival of the patient. However, bacterial analysis in ascitic fluid is currently based on culture methods, which are time-consuming and laborious. We report here the application of a photonic interferometer biosensor based on a bimodal waveguide (BiMW) for the rapid and label-free detection of bacteria directly in ascitic fluid. The device consists of a straight waveguide in which two modes of the same polarization interfere while interacting with the external medium through their evanescent fields. A bimolecular event occurring on the sensor area of the device (e.g. capturing bacteria) will differently affect each light mode, inducing a variation in the phase of the light exiting at the output of the waveguide. In this work, we demonstrate the quantitative detection of Bacillus cereus in buffer medium and Escherichia coli in undiluted ascitic fluid from cirrhotic patients. In the case of Bacillus cereus detection, the device was able to specifically detect bacteria at relevant concentrations in 12.5min and in the case of Escherichia coli detection, the analysis time was 25min. Extrapolation of the data demonstrated that the detection limits of the biosensor could reach few bacteria per milliliter. Based on the results obtained, we consider that the BiMW biosensor is positioned as a promising new clinical tool for user-friendly, cost-effective and real-time microbiological analysis.

Diagnostic and prognostic role of procalcitonin (PCT) and MR-pro-Adrenomedullin (MR-proADM) in bacterial infections.

Rapid diagnosis of bacterial infections is crucial for adequate antibiotic treatment. Serum molecules such as Procalcitonin (PCT) have been used as biomarkers of infection. Recently, the mid-regional pro-Adrenomedullin (MR-proADM) has been evaluated in combination with PCT for sepsis diagnosis. The diagnostic role of PCT and MR-proADM both in sepsis and in localized infections together with their contribution to effective antibiotic therapy has been evaluated. One hundred and eighty-two patients with bacterial infection has been enrolled: PCT and MR-proADM were measured at admission (T=0), at 12-24h (T=1) and in the third or fifth day of antibiotic therapy (T=3-5). ROC curve (receiver operating characteristic) and post-test probability were calculated. MR-proADM increased with the severity of the infection. PCT resulted significantly higher in sepsis than localized infection. After antibiotic therapy, PCT significantly decreased in localized respiratory infections and in sepsis, while MR-proADM decreased significantly after antibiotic therapy only in patients with severe sepsis/septic shock. The threshold values of PCT and MR-proADM were >0.1ng/mL and >0.8nmol/L, respectively. The combined use of PCT and MR-proADM increased the post-test probability of the diagnosis of bacterial infections compared to PCT alone. In conclusion, PCT and MR-proADM combination improves the diagnosis of bacterial infection and contribute to prognosis and antibiotic therapy effectiveness.

Rapid antimicrobial susceptibility testing with electrokinetics enhanced biosensors for diagnosis of acute bacterial infections.

Rapid pathogen detection and antimicrobial susceptibility testing (AST) are required in diagnosis of acute bacterial infections to determine the appropriate antibiotic treatment. Molecular approaches for AST are often based on the detection of known antibiotic resistance genes. Phenotypic culture analysis requires several days from sample collection to result reporting. Toward rapid diagnosis of bacterial infection in non-traditional healthcare settings, we have developed a rapid AST approach that combines phenotypic culture of bacterial pathogens in physiological samples and electrochemical sensing of bacterial 16S rRNA. The assay determines the susceptibility of pathogens by detecting bacterial growth under various antibiotic conditions. AC electrokinetic fluid motion and Joule heating induced temperature elevation are optimized to enhance the sensor signal and minimize the matrix effect, which improve the overall sensitivity of the assay. The electrokinetics enhanced biosensor directly detects the bacterial pathogens in blood culture without prior purification. Rapid determination of the antibiotic resistance profile of Escherichia coli clinical isolates is demonstrated.

Titania nanotube-modified screen printed carbon electrodes enhance the sensitivity in the electrochemical detection of proteins

The use of titania nanotubes (TiO2-NT) as the working electrode provides a substantial improvement in the electrochemical detection of proteins. A biosensor designed using this strategy provided a robust method to detect protein samples at very low concentrations (Cprotein ca 1ng/μl). Reproducible measurements on protein samples at this concentration (Ip,a of 80+1.2μA) could be achieved using a sample volume of ca 30μl. We demonstrate the feasibility of this strategy for the accurate detection of penicillin binding protein, PBP2a, a marker for methicillin resistant Staphylococcus aureus (MRSA). The selectivity and efficiency of this sensor were also validated using other diverse protein preparations such as a recombinant protein tyrosine phosphatase (PTP10D) and bovine serum albumin (BSA). This electrochemical method also presents a substantial improvement in the time taken (few minutes) when compared to conventional enzyme-linked immunosorbent assay (ELISA) protocols. It is envisaged that this sensor could substantially aid in the rapid diagnosis of bacterial infections in resource strapped environments.

An integrated microfluidic device utilizing vancomycin conjugated magnetic beads and nanogold-labeled specific nucleotide probes for rapid pathogen diagnosis

A PCR-free assay for rapid pathogen diagnosis was implemented on an integrated microfluidic system in this study. Vancomycin-conjugated magnetic beads were used to capture multiple strains of bacteria and nucleotide probes labeled gold nanoparticles were used to specify and detect a specific strain by hybridization-induced color change. The assay was entirely automated within an integrated microfluidic device that was composed of suction-type micropumps, microvalves, microchannels, and microchambers that fabricated by microfluidic technology. Multiple strains of bacteria could be captured simultaneously by vancomycin-conjugated magnetic beads, with capturing efficiency exceeding 80%. Subsequently, sensitive and strain-specific detection against target bacteria could be achieved by using nanogold labeled specific nucleotide probes. The limit of detection of 102CFU bacteria was achieved. Importantly, nucleic acid amplification was not involved in the diagnostic procedures; the entire analytic process required only 25min. The developed platform may provide a promising tool for rapid diagnosis of bacterial infections. From the Clinical EditorIn this novel study, a PCR-free pathogen detection method is demonstrated. After vancomycin-conjugated magnetic beads captured bacteria, nucleotide probes-labeled gold nanoparticles were employed to specify and detect specific strains via hybridization-induced color change. Multiple strains of bacteria could be captured simultaneously with an efficiency exceeding 80%, enabling the detection of as low as 102 CFU of bacteria.

A new single‐step quantitative pathogen detection system: Template‐tagging followed by multiplex asymmetric PCR using common primers and CE‐SSCP

Rapid diagnosis of bacterial infection is important for patient management and appropriate therapy during the early phase of bacteria-induced disease. Among the existing techniques for identifying microbial, CE-SSCP combined with 16S ribosomal RNA gene-specific PCR has the benefits of excellent sensitivity, resolution, and reproducibility. However, even though CE-SSCP can separate PCR products with high-resolution, multiplex detection and quantification are complicated by primer-dimer formation and non-specific amplification. Here, we describe a novel technique for multiplex detection and quantification of pathogens by template-tagging followed by multiplex asymmetric PCR and subsequent CE-SSCP. More specifically, we reverse transcribed 16S ribosomal RNAs from seven septicemia-inducing pathogens, tagged the templates with common end sequences, and amplified them using common primers. The resulting amplicons could be successfully separated by CE-SSCP and quantified by comparison to an internal standard. This method yielded results that illustrate the potential of this system for diagnosing infectious disease.

Genotyping DNA chip for the simultaneous assessment of antibiotic resistance and pathogenic potential of extraintestinal pathogenic Escherichia coli

Urinary tract infections (UTIs) are among the most frequently occurring infections and are mostly caused by extraintestinal pathogenic Escherichia coli. DNA microarrays are potent molecular diagnostic tools for rapid diagnosis of bacterial infections with high relevance for UTIs. In this study, we present the integration and application of two DNA chip modules for the simultaneous detection of single nucleotide polymorphisms in gyrA (quinolone resistance) and fimH (increased adhesion to urinary tract epithelium). The performance of the combined diagnostic chip was assessed by genotyping 140 E. coli strains. Resistance-causing mutations could only be identified in UTI isolates. A complete genotyping assay could be performed in <4 h after DNA extraction. Together with the excellent genotyping results, this constitutes a competitive alternative as a standard tool for routine clinical diagnostics.

Rapid Diagnosis of Bacterial Infections Using Technologies Based on Nucleic Acid Detection

Bacterial identification and antibiotic susceptibility testing methods used in clinical microbiology laboratories generally require at least two days. This long delay often forces physicians to treat patients presumptively with broad‐spectrum antibiotics. Novel diagnostic tests based on the detection of nucleic acids (DNA or RNA) offer a great potential for the rapid (approximately 1 h) diagnosis of bacterial infections. The present article reviews various aspects of the development and validation of nucleic acid‐based assays suitable for the detection and identification of bacteria as well as for the detection of associated antibiotic resistance genes. The potential of these assays for routine use in clinical microbiology laboratories is also discussed.

Novel Approaches to the Prevention and Therapy of Neonatal Bacterial Sepsis

A variety of adjunctive treatments have been shown to offer potential benefits for neonates with sepsis. Most are not available clinically and those that are available still should be considered experimental or limited in their use; however, these efforts are far from complete and should continue to evolve in the coming years. Efforts toward the rapid diagnosis of bacterial infections are a necessary component in the eventual implementation of these potential novel strategies. As a better understanding of the intricate mechanisms of neonatal sepsis is developed, it will be possible to provide patients with an increasingly effective array of treatment and prevention strategies.