| dc.description.abstract | According to a recent investigation by the World Health Organization (WHO) in 2021, every
year, nearly 400 million people worldwide are affected by dengue virus (DENV), a mosquitoborne illness caused by DENV (WHO, 2021). Particularly, Vietnam is one of the most seriously
affected countries recorded a high number of DENV infections (ECDC, 2021). In the situation of
lacking vaccination, and therapy drugs of DENV infection, the timely and accurate diagnosis is
crucial to the general public's health. The current gold standard for DENV infection detection and
serotyping is reverse-transcriptase (RT) quantitative polymerase chain reaction (qPCR) tests.
However, RT-qPCR must be performed in a laboratory setting using specialist equipment and
personnel with the appropriate training. Therefore, PCR-based methods have limitations in solving
the problem of rapid diagnosis as well as deployment in low-middle income countries (LMICs)
where DENV infection is most serious.
Isothermal nucleic acid amplification techniques have recently surfaced as a substitute for
traditional deoxyribonucleic acid (DNA) amplification methods, with the goal of simplifying
molecular diagnostics. One of them, loop-mediated isothermal amplification (LAMP) assay, is a
sensitive, specific, simple, rapid, and cheap molecular method which does not necessitate
thermocyclers, so it could potentially be used outside of laboratories or downsized (Mori et al.,
2021). Due to LAMP's superior features, it is a promising alternative to other molecular
amplification techniques, especially for applications in rapid diagnosis, and point-of-care (POC)
testing of DENV infection. But the superior properties of LAMP depend on the use of multiple
primers and Bacillus stearothermophilus (Bst) DNA polymerase which leads to dimer formation.
As a result, the potential of false-positive results and low sensitivity resulting from nonspecific
and nontemplate LAMP amplification may reduce test reliability.
In this project, a protocol for a LAMP assay to identify the DENV-NS1 gene for Dengue
Virus detection was developed. Moreover, to improve the performance of the LAMP reaction, this
assay has applied two enhancement strategies, including the prehybridization-induced enhancement (PIE) strategy and organic additives. The enhanced LAMP reaction results after
incubation at 65°C for 30 minutes and reaches a limit of detection ~46 copies/µl. | en_US |
