LIBIRAN, AIRA MICHAELLA G.

DENGUE VIRUS (DENV)

HOW IS IT IDENTIFIED IN THE LABORATORY?

 Dengue Virus Antigen Detection

o NS1 Detection - It detect the non-structural protein NS1 of dengue virus. This protein is secreted into the
blood during dengue infection. This test has been developed for use in serum. Most of these tests use
synthetically labeled antibodies to detect dengue NS1 protein. NS1 is detectable during the acute phase of
dengue virus infections. NS1 tests can be as sensitive as molecular tests during the first 0-7 days of symptoms.
After day 7, NS1 tests are not recommended.

 Serologic Tests for Dengue Virus

o IgM Antibody Capture Enzyme-Linked Immunosorbent Assay (MAC-ELISA) - The dengue MAC-ELISA is used
for the qualitative detection of dengue virus IgM antibodies. The MAC-ELISA is based on capturing human IgM
antibodies on a microtiter plate using anti-human-IgM antibody followed by the addition of dengue virus
antigens. The antigens used for this assay are derived from the envelope proteins of the four dengue virus
serotypes (DENV-1-4). Combined testing with a nucleic acid amplification test (NAAT) and MAC-ELISA usually
provides a diagnostic result during the first 1-7 days of illness. A convalescent phase specimen is needed to
make a diagnosis of dengue virus infection when results are negative on both tests from the acute specimen.
o Plaque Reduction Neutralization Test (PRNT) – PRNT is used to more precisely determine the cause of
infection in IgM positive patients for whom a specific diagnosis is clinically or epidemiologically important
(e.g., to rule out other flaviviruses such as Zika or yellow fever).
PRNT is used to confirm the infecting virus in a dengue or Zika virus IgM-positive specimen and can in some
cases determine the infecting dengue virus serotype.

 Tissue Tests for Dengue Virus

o Nucleic Acid Amplification Test (NAAT) for Fixed Tissue Specimens - A NAAT, like real-time reverse
transcription polymerase chain reaction (rRT-PCR) or in situ hybridization, detects dengue virus RNA. NAATs
are less sensitive after day 7 of illness.

 Molecular Tests for Dengue Virus

o Nucleic Acid Amplification Test (NAAT) - A NAAT is a generic term referring to molecular tests used to detect
viral genomic material. NAAT assays are the preferred method of diagnosis, because they can provide
confirmed evidence of infection. For symptomatic patients during the first 1-7 days of illness, any serum
sample should be tested with a NAAT and for IgM antibody since both tests can be performed in serum.
Performing both tests can detect more cases than performing just one test. After day 7 of illness, few cases
can be detected by NAAT.

 Hematological Tests - Standard hematological parameters such as platelet count and hematocrit are important
and are part of the biological diagnosis of dengue infection. Therefore, they should be closely monitored.
Thrombocytopenia, a drop in platelet count below 100 000 per µl, may be occasionally observed in dengue fever
but is a constant feature in DHF. Thrombocytopenia is usually found between the third and eighth day of illness
often before or simultaneously with changes in hematocrit. Hemoconcentration with an increase in the
 hematocrit of 20% or more (for the same patient or for a patient of the same age and sex) is considered to be a
definitive evidence of increased vascular permeability and plasma leakage.

HOW IS IT INDENTIFIED UNDER MICROSCOPE?

Dengue virus-like particles in platelets isolated from confirmed dengue patients.

a) Dengue virus-like particles were observed inside vesicle compartment (red arrow) and a particle appeared to be on its
way budding out into the vesicle (blue arrow). b) A single fuzzy virus-like particle was released from platelet (red arrow
head)

Structure of Dengue Virus

WHAT IS THE PROCEDURE BEING IDENTIFIED OR CULTIVATED?

PHYSICAL PROPERTIES

DISEASES CAUSED

DIFFERENT MANAGEMENT IN PREVENTING THE SPREAD OF THE VIRUS