Helicase-dependent isothermal amplification: a novel tool in the development of molecular-based analytical systems for rapid pathogen detection

• Published in: Analytical and bioanalytical chemistry 2018-01, Vol.410 (3), p.679-693
• Main Authors: Barreda-García, Susana, Miranda-Castro, Rebeca, de-los-Santos-Álvarez, Noemí, Miranda-Ordieres, Arturo J., Lobo-Castañón, María Jesús
• Format: Article
• Language: English
• Subjects: ABCs 16th Anniversary; Acids; Amplification; Analytical Chemistry; Animals; Bacteria - isolation & purification; Bacterial Infections - diagnosis; Bacterial Infections - microbiology; Biochemistry; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; DNA helicase; DNA Helicases - chemistry; DNA polymerase; DNA, Bacterial - analysis; DNA, Viral - analysis; E coli; Electrochemical Techniques - instrumentation; Electrochemical Techniques - methods; Environmental monitoring; Environmental Monitoring - instrumentation; Environmental Monitoring - methods; Environmental testing; Food safety; Food Science; Hazard Analysis and Critical Control Points - methods; Health aspects; Humans; Infectious diseases; Laboratories; Laboratory Medicine; Microfluidic Analytical Techniques - instrumentation; Microfluidic Analytical Techniques - methods; Microfluidics; Microorganisms; Monitoring/Environmental Analysis; Nucleic Acid Amplification Techniques - instrumentation; Nucleic Acid Amplification Techniques - methods; Nucleic acids; Pathogens; Proteins; Public health; Review; Sample preparation; Sensitivity analysis; Systems analysis; Technology application; Temperature; Thermal cycling; Virus Diseases - diagnosis; Virus Diseases - virology; Viruses - isolation & purification
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-017-0620-3

Highly sensitive testing of nucleic acids is essential to improve the detection of pathogens, which pose a major threat for public health worldwide. Currently available molecular assays, mainly based on PCR, have a limited utility in point-of-need control or resource-limited settings. Consequently, there is a strong interest in developing cost-effective, robust, and portable platforms for early detection of these harmful microorganisms. Since its description in 2004, isothermal helicase-dependent amplification (HDA) has been successfully applied in the development of novel molecular-based technologies for rapid, sensitive, and selective detection of viruses and bacteria. In this review, we highlight relevant analytical systems using this simple nucleic acid amplification methodology that takes place at a constant temperature and that is readily compatible with microfluidic technologies. Different strategies for monitoring HDA amplification products are described. In addition, we present technological advances for integrating sample preparation, HDA amplification, and detection. Future perspectives and challenges toward point-of-need use not only for clinical diagnosis but also in food safety testing and environmental monitoring are also discussed. Graphical Abstract Expanding the analytical toolbox for the detection of DNA sequences specific of pathogens with isothermal helicase dependent amplification (HDA)