Abstract

A trend towards the abandonment of obtaining pure culture isolates in frontline laboratories is at a crossroads with the ability of public health agencies to perform their basic mandate of foodborne disease surveillance and response. The implementation of culture-independent diagnostic tests (CIDTs) including nucleic acid and antigen-based assays for acute gastroenteritis is leaving public health agencies without laboratory evidence to link clinical cases to each other and to food or environmental substances. This limits the efficacy of public health epidemiology and surveillance as well as outbreak detection and investigation. Foodborne outbreaks have the potential to remain undetected or have insufficient evidence to support source attribution and may inadvertently increase the incidence of foodborne diseases. Next-generation sequencing of pure culture isolates in clinical microbiology laboratories has the potential to revolutionize the fields of food safety and public health. Metagenomics and other ‘omics’ disciplines could provide the solution to a cultureless future in clinical microbiology, food safety and public health. Data mining of information obtained from metagenomics assays can be particularly useful for the identification of clinical causative agents or foodborne contamination, detection of AMR and/or virulence factors, in addition to providing high-resolution subtyping data. Thus, metagenomics assays may provide a universal test for clinical diagnostics, foodborne pathogen detection, subtyping and investigation. This information has the potential to reform the field of enteric disease diagnostics and surveillance and also infectious diseases as a whole. The aim of this review will be to present the current state of CIDTs in diagnostic and public health laboratories as they relate to foodborne illness and food safety. Moreover, we will also discuss the diagnostic and subtyping utility and concomitant bias limitations of metagenomics and comparable detection techniques in clinical microbiology, food and public health laboratories. Early advances in the discipline of metagenomics, however, have indicated noteworthy challenges. Through forthcoming improvements in sequencing technology and analytical pipelines among others, we anticipate that within the next decade, detection and characterization of pathogens via metagenomics-based workflows will be implemented in routine usage in diagnostic and public health laboratories.

Highlights

  • The incidence and impact of foodborne illness constitutes a significant global issue to public health

  • Clinical microbiology laboratories could perform reflex culturing of biological specimens that test positive via culture-independent diagnostic tests (CIDTs) such that positive isolates can still be submitted to public health laboratories for subtyping or other culture-based tests such as WGS (Cronquist et al, 2012)

  • The prospective use of diagnostic metagenomics and comparable techniques offers an assumption-free workflow creating the ability to detect any and all pathogens from various biological specimens or food products

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Summary

INTRODUCTION

The incidence and impact of foodborne illness constitutes a significant global issue to public health. Pathogen detection is increasingly contingent upon the analytic application of CIDTs, which include nucleic acid (e.g., PCR) and antigen-based tests (e.g., ELISA) among others. The use of nucleic acid-based assays to identify enteric pathogens presents with several advantages over culturedependent tests in clinical and food settings. Their use allows for high levels of sensitivity and specificity and offers an added benefit in their ability to detect toxin-producing genes or other important biomarkers. A growing number of US FDA – approved syndromic panels for multiple pathogen detection in addition to laboratorydeveloped tests have facilitated the upsurge of CIDT application for acute gastroenteritis in clinical laboratories These panels employ PCR to detect unique DNA sequences to enable identification of enteric pathogens. Multiplex panels are highly sensitive (90–100%) and specific (98%) (Buchan et al, 2013; Navidad et al, 2013; Onori et al, 2014; Buss et al, 2015; Harrington et al, 2015)

Whole Genome Sequencing
Diagnostic Metagenomics and Comparable Detection Techniques
Metagenomics and Comparable Detection Techniques in Food Safety
Technical expertise
Suggested sequencing approach
Functional profiling
Technical Challenges of Metagenomics Sequencing and Analysis
Pathogen Surveillance and Subtyping
Genomic Epidemiology
Metagenomics is Capable of Providing Informative Subtyping Data
Changing Trends in Public Health Surveillance
Whole Genome Sequencing Antimicrobial Resistance Gene Detection
Metagenomics Antimicrobial Resistance Gene Detection
Current Condition of Antimicrobial Resistance Surveillance
FUTURE CHALLENGES OF METAGENOMICS IN DIAGNOSTIC AND PUBLIC HEALTH SETTINGS
Findings
CONCLUSION
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