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2015 ; 5
(ä): 17925
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A novel multiplex isothermal amplification method for rapid detection and
identification of viruses
#MMPMID26643761
Nyan DC
; Swinson KL
Sci Rep
2015[Dec]; 5
(ä): 17925
PMID26643761
show ga
A rapid multiplex isothermal amplification assay has been developed for detection
and identification of multiple blood-borne viruses that infect millions of people
world-wide. These infections may lead to chronic diseases or death if not
diagnosed and treated in a timely manner. Sets of virus-specific oligonucleotides
and oligofluorophores were designed and used in a reverse-transcription
loop-mediated multiplexed isothermal amplification reaction for detection and gel
electrophoretic identification of human Immunodeficiency virus (HIV), hepatitis-B
virus (HBV), hepatitis-C virus (HCV), hepatitis-E virus (HEV), dengue virus
(DENV), and West Nile (WNV) virus infection in blood plasma. Amplification was
catalyzed with two thermostable enzymes for 30-60 minutes under isothermal
condition, utilizing a simple digital heat source. Electrophoretic analysis of
amplified products demonstrated simultaneous detection of 6 viruses that were
distinctly identified by unique ladder-like banding patterns. Naked-eye
fluorescent visualization of amplicons revealed intensely fluorescing products
that indicated positive detection. The test demonstrated a 97% sensitivity and a
100% specificity, with no cross-reaction with other viruses observed. This
portable detection tool may have clinical and field utility in the developing and
developed world settings. This may enable rapid diagnosis and identification of
viruses for targeted therapeutic intervention and prevention of disease
transmission.