Challenges in Direct Detection of Flaviviruses: A Review

Abstract

Arthropods transmit arboviruses via mosquito and tick bites to humans and other animals. The genus flavivirus, which causes diseases, sequelae, and thousands of deaths, mainly in developing and underdeveloped countries, is among the arboviruses of interest to public health. Given the importance of early and accurate diagnosis, this review analyzes the methods of direct detection of flaviviruses, such as reverse transcription loop-mediated isothermal amplification, microfluidics, localized surface plasmon resonance, and surface-enhanced Raman scattering, and presents the advantages, disadvantages, and detection limits identified in studies reported in the literature for each methodology. Among the different methods available, it is essential to balance four fundamental indicators to determine the ideal test: good sensitivity, high specificity, low false positive rate, and rapid results. Among the methods analyzed, reverse transcription loop-mediated isothermal amplification stands out, owing to result availability within a few minutes, with good sensitivity and specificity; in addition, it is the best-characterized methodology.

Tick-borne viruses and their risk to public health in the Caribbean: Spotlight on bats as reservoirs in Cuba

“…Flaviviruses, belonging to the Flaviviridae family, are transmitted by hematophagous mosquitoes or ticks and have the potential to cause epidemics with significant morbidity and mortality, making them a global health problem. Flaviviruses of medical importance transmitted by mosquitoes include West Nile virus (WNV), Japanese encephalitis virus [ 33 ], Yellow fever virus (YFV), Dengue virus (DENV), and Zika virus (ZIKV) [ 34 ]. While tick-transmitted flaviviruses have historically garnered less attention due to their comparatively lower incidence in human infections and the greater medical impact of mosquito-transmitted counterparts, the emergence of the Severe Fever with Thrombocytopenia Syndrome virus (SFTSV) in China in 2011 has shifted focus.…”

CRISPR-Cas: ‘The Multipurpose Molecular Tool’ for Gene Therapy and Diagnosis

“…The ‘heating unextracted diagnostic samples to obliterate nucleases’ (HUDSON) methodology was developed to identify viral genetic material from body fluids, such as urine, blood and its isolates, and saliva, making the SHERLOCK procedure even more efficient [ 53 ]. The HUDSON protocol is used for conserved regions within the genetic material of viruses identified using universal-flavivirus recombinase polymerase amplification (RPA) primers [ 54 ]. The SHERLOCK and HUDSON protocols can be applied to any virus, but previous testing focused on the diagnosis of flaviviruses, such as the Zika, Dengue, West Nile, and yellow fever viruses [ 2 , 19 ].…”

In Situ Colorimetric LAMP Based on One-Step Modified Filter Paper to Screen Human Papillomavirus (HPV)16/18 from Clinical Samples

“…34−36 For colorimetric detection of the LAMP assay, pH-sensitive dyes and metal ion indicators have been reported as color indicators in the literature. 32,37,38 The result can be obtained quickly by observing the color change with the naked eye, which can further reduce the dependence on instruments. 39 Although the process of HPV genotype identification could be greatly simplified although a visual LAMP assay, pretreatment of raw samples still relies on auxiliary equipment, such as centrifuges, for cell enrichment, separation, lysis and nucleic acid extraction, which increases the dependence of wholeprocess detection on equipment and reduces the potential for popularization and application.…”

“…Compared with PCR-based detection, isothermal amplification methods are better suited for on-site rapid detection, with the use of a much simpler and less expensive heating instrument to run the nucleic acid assay. − These methods include LAMP, − nucleic acid sequence-based amplification (NASBA), recombinase polymerase amplification (RPA), , helicase-dependent amplification (HDA), and strand displacement amplification (SDA) . Among these isothermal techniques, LAMP has been most widely researched for POC applications. ,,, The LAMP assay can be performed by examining the turbidity or color change of the reaction solution. − For colorimetric detection of the LAMP assay, pH-sensitive dyes and metal ion indicators have been reported as color indicators in the literature. ,, The result can be obtained quickly by observing the color change with the naked eye, which can further reduce the dependence on instruments …”