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Amplification of Near Full-length HIV-1 Proviruses for Next-Generation Sequencing

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Published on: October 16, 2018

Methods in virus diagnostics: from ELISA to next generation sequencing.

Neil Boonham¹, Jan Kreuze², Stephan Winter³

¹The Food & Environment Research Agency (FERA), Sand Hutton, York YO41 1LZ, UK.

|December 24, 2013

Summary

This summary is machine-generated.

New plant virus diagnostic tools must be easy to use and establish, not just offer performance benefits over enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (PCR). Human factors and stakeholder engagement are key to adopting novel virus detection methods.

Keywords:

Field testing LAMP assays Molecular diagnostics Multiplex, Luminex Next generation sequencing Real-time PCR

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Isolation and Genome Analysis of Single Virions using 'Single Virus Genomics'

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Published on: May 26, 2013

Area of Science:

Plant virology
Diagnostic technology development
Biotechnology

Background:

Routine plant virus detection relies on established methods like enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (PCR).
Despite advancements, newer diagnostic technologies face low adoption rates in routine laboratory settings.
Understanding the characteristics of successful routine diagnostic tools is crucial for evaluating new methods.

Purpose of the Study:

To analyze the factors influencing the adoption of new plant virus diagnostic technologies.
To identify the essential attributes of routine diagnostic tools by examining established methods.
To explore the potential and challenges of emerging diagnostic approaches.

Main Methods:

Comparative analysis of established diagnostic methods (ELISA, real-time PCR) to identify key success factors.
Review of emerging plant virus detection technologies, including loop-mediated isothermal amplification (LAMP), Luminex bead arrays, and next-generation sequencing (NGS).
Assessment of field-based diagnostics like Lateral Flow Devices (LFDs) and their limitations.

Main Results:

Established methods like ELISA and real-time PCR offer accessibility, ease of use, and robustness.
Emerging technologies focus on field applicability (LAMP), multiplexing (Luminex), and virus discovery (NGS).
Barriers to adopting field-based technologies are often behavioral, not performance-related, necessitating new collaborations.

Conclusions:

Future routine diagnostics must offer advantages over current methods and be user-friendly for widespread adoption.
Next-generation sequencing (NGS) shows promise for convergence of discovery and multiplexing, but data analysis expertise is critical.
Human factors, including user behavior, laboratory practices, and policy, alongside technical performance, will ultimately determine the success of new diagnostic tools.