Rapid discrimination of

Malaria and Dengue

Infected Patients Sera using
Raman Spectroscopy
• Present diagnosis relies on the identification of Plasmodium,
causative agent of malaria in blood and the “gold standard” being
histopathological analysis of Giemsa stained blood-smears.
• Alternative routes:

MRDT kit detects persisting antigens such as histidine-rich protein II (HRPII) and lactate dehydrogenase for
falciparum and vivax infection respectively.

• Studies show that missed or delayed diagnosis of malaria in 59-71%

of imported cases or in non-endemic regions, for instance, dengue
endemic area adds-on to morbid and fatal outcomes.
• Global escalation of dengue incidence, approximately 3.9 billion cases
in 128 countries have worsened the situation.
• Dengue diagnosis on serological tests:

• most commonly used combination:

1. the requirement of acute sample, expertise and appropriate
facilities, time-consuming (sometimes more than 1 week),
expensive.
2. cannot differentiate between primary and secondary infection, IgM
levels at times are below recognition due to secondary infections
and may lead to misdiagnosis.
3. physicians in endemic areas sometimes only use positive tourniquet
test.
4. Dengue RDT kit resulted to “False positive”
• Considering the diagnostic limitations, this study aims to develop
diagnostic-methods for simultaneous discrimination of malaria and
dengue infection using .

• Timecourse study of mice plasma infected with Plasmodium clearly

indicated that RS could accurately detect early stages of infection,
with parasitemia levels as low as 0.2%, which is typically difficult to be
detected by existing methods.
 on human subjects:

Blood sample are obtained from 37 malaria and 39 dengue patients,

and 54 Healthy Cases.
Frozen serum (30 μl) of malaria, dengue, and HC following passive
thawing was individually placed on CaF2 window to record spectra
using HE-785 commercial RS (Jobin-Yvon-Horiba, France) with fiber-
optic microprobe.
• Total bilirubin, urea, liver enzymes such as alanine aminotransferase
(ALT), aspartate transaminase (AST), and alkaline phosphatase (ALP)
were significantly elevated in both diseased groups compared to HC
(Table S1), suggesting discrimination of malaria and dengue is difficult
based on the clinicopathological features.
• RS is more robust than RDTs or PCR that employs only one or two
specific antigens/antibodies for disease diagnosis. Besides, RS has
been successfully employed in diagnostics, prognostics or as a tool for
evaluating new therapies for several diseases including cancer,
urology and andrology, asthma, tuberculosis and bone
demineralization.
• The RS model developed in the study is very reliable, accurate, time-
efficient and requires minimal sample-processing.
• One of the drawbacks of the study is small sample size, however the
limitation was compensated by using complementary MS-based
approach to validate the findings.
• The further in-depth study may lead to the identification of a novel
candidate for drug targets and the study could be translated to the
field-settings particularly in malaria and dengue-endemic regions to
screen larger patient cohorts.
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