SUMMARY

“Urine-Saliva Stick as Inovation of Antigen NS1 and Antibody IgG Dengue

Virus Infection using Immunochoromatography”

Yuvita Dian Damayanti (142210101025)

Indonesia is a tropical country located in Southeast Asia, affecting the

breeding of disease vectors, including mosquito. Based on data from the World
Health Organization (WHO), Indonesia is a country with the most cases of dengue
fever in Southeast Asia. The prevalence of dengue infection has reached 5 to 100
million people every year and more than 2.5 billion population which is about 40% of
the world's population at risk of the disease.

Dengue hemorrhagic fever (DHF) is a disease caused by dengue virus which

is included in the family Flaviridae with humans as the hosts. This disease is caused
by the bite of Aedes aegypti and Aedes albopictus infected by the virus. This disease
is dangerous because it can lead to death in a short duration. Until now, the therapy
used to treat this disease can not be specific. Thus, early careful and quick detection is
necessary to prevent dengue virus infection that eventually became worse dengue
disease.

Nowadays, there are rapid diagnostic tests for dengue infection. Some existing
diagnostic tests are NS1 antigen using PCR technique and IgM and IgG tests by
ELISA. But, most of them still use whole blood, serum, or plasma as sample, so they
are not appropriate for children or patients who have needle phobia. Therefore, non-
invasive methods are needed to perform an early diagnosis of dengue virus infection
and the patients can do the sample collection methods by themselves, eg, using a
sample of urine or saliva. Combination detection of the NS1 antigen and IgG
antibody is used to determine the primary or secondary infection of the patients.

NS1 antigen is a glycoprotein produced by a Flavivirus which is useful in the

process of viral replication and life. This antigen will appear on the first day after the
onset of fever appears, so the NS1 antigen detection can use for early detection of
dengue virus. Meanwhile, IgG antibody can be used as a marker of dengue virus
infection to distinguish primary and secondary infection using urine and saliva
samples. IgG antibody in urine and saliva samples will appear starting on the fourth
to fifth day in secondary infection, whereas in primary infection, IgG antibody will
not be detected in urine and saliva samples until the seventh day.

Immunochromatography consist of nitrocellulose membrane as a capillary

membrane, antibody or antigen-specific is fixed in T line (test line), a recombinant
protein or another antibody fixed at C line (line of control), and the dye labeled
antibody to be identified visually. In the membrane nitrocellulose anti-NS1 antibodies
that will form a complex with the NS1 antigen contained in urine and saliva samples.
The complex will move towards the test area (line 'T') mobilized by the others anti-
dengue NS1 antibody causing changes color to burgundy. If the color does not
appear, the interpretation of the results is the sample does not contain NS1 antigen.
Burgundy 'C' line meaning the control on the tool shows the correct procedure,
otherwise if the color does not appear, the test procedure should be repeated.
Similarly, in the nitrocellulose membrane containing antigen-specific IgG for
detecting IgG antibody in samples.

A negative result obtained when only color appears on the both ‘C’ lines. This
indicates that there is no NS1 antigen and IgG antibody in the sample. Secondary
infection dengue virus obtained in two conditions if color appears on the both ‘C’
lines, NS1 line and IgG line or color appears on the both ‘C’ lines and IgG line.
Primary infection dengue virus obtained if color appears on the both ‘C’ lines and
NS1 line. The results are invalid, if the color appears on the NS1 line or IgG alone,
NS1 alone, IgG alone, or no color appears on all lines. It indicates that the procedure
is incorrect or the tool is damage.

The advantages using Urine-Saliva Stick are the method is fast (about 15-20
minutes), non-invasive, for early detection of dengue virus infection, to distinguish
between primary and secondary infection, easy sampling, suitable for children or
adults who have needle phobia. However the accuracy, sensitivity, and specificity of
this tool is not yet known, thus requiring further research.