1. Diagnosis of Human
Filariasis
ubio sensit
Filariasis
Antibody
Test

2. lymphatic filariasis
 Long known disease which only in a minority of cases
results in severe elephantiasis
 120 million people infected of which 40 million show
symptoms (1996)
 Caused by three species of filaria: Wucheria bancrofti,
Brugia malayi, B. timori

3. Lymphatic Filariasis: Life Cycle

4. Filaria and filariasis
Tissue dwelling nematodes with adults and L1 present in the
human host
Arthropode vector which takes up L1 and transmits L3
Onchocerciasis, Lymphatic filariasis, (Loiasis)

5. Are endosymbionts involved in filarial
pathogenesis?
 Wolbachia bacteria are found as
intracellular endosymbionts in
the hypodermal lateral chord
and the uterus of many parasitic
nematodes (here Brugia malayi)
 The bacteria seem important
for nematode development as
antibiotic treatment results in
sterility (and maybe even death)
of the adults

6. Are endosymbionts involved in filarial
pathogenesis?
 Sections through onchocercomas in untreated
(upper) and antibiotic treated patients (11
months after a 6 weeks course of doxycyclin)
 Note that Wolbachia detected by red stain are
absent in the treated patient
 In addition the treated worms show no healthy
eggs or embryos
 This suggest that antibiotics might provide
new therapeutic approaches for the treatment
of filariasis
 There are also studies that suggest that the
bacterial antigens might be responsible for the
strong inflammation and hence pathology

7. lymphatic filariasis
Wucheria bancrofti
Brugia malayi

8. lymphatic filariasis
 Adult worms (macrofilariae)
live in the lymphatic vessels and
lymph nodes of the lower body
half
 Females are ovoviviparous
producing L1 larvae still in the
egg membrane (the sheath)

9. lymphatic filariasis
 L1 lavae or microfilariae are
swept into the blood stream and
circulate
 W.b. microfilariae are sheated
and the nuclei do not reach the
tip of the tail
 Microfilariae remain viable and
infective for several months

10. lymphatic filariasis
 Microfilaria show diurnal rhythm
 Wucheria and Brugia microfilariae
are found in the peripheral blood
during night time whereas Loa is
found during the day hours
 This is phenomenon is not linked to
egg production but to the behavior
of the MF
 The benefit (of absence from the
peripheral blood during the day is
unclear)

11. lymphatic filariasis
 Mircofilariae are taken up by
mosquitoes with the blood meal
 Broad spectrum of night active
vectors (Culex, Aedes,
Anopheles, Mansonia)
 Microfilaria leave sheath in the
mosquito midgut, penetrate the
Culex quiquefasciatus
midgut wall and migrate to the
flight muscle where they molt
twice (the develop intra-
cellularly)
W. bancrofti L1 in mosquito flight muscle

12. lymphatic filariasis
 L3 migrate through hemolymph until they find the labium,
which they penetrate when they sense that the mosquito is
feeding
 They move onto the skin and into the wound puncture
 Development in mosquito takes about two weeks

13. lymphatic filariasis
 Maturing larvae and adults provoke
strong inflammatory reaction
 Acute symptoms are painful
lymphnode and lymphchannel
inflammation and swelling which is
often accompanied by fever
 Brugia infection is very similar to
Wucheria
 Acute reactions are more pronounced
(e.g. formation of abscesses)
 Elephantiasis tends to affect arms
instead of legs

14. lymphatic filariasis
 Progessive chronic disease can
lead to wide spread fibrosis and
damage of lymphatic vessels,
which can result in rupture and
discharge of lymph into the
urinary system (chyluria) or the
scrotum

15. lymphatic filariasis
 In men chronic infection often
results in hydrocele, a painful
swelling of the scrotum due to
blockage and inflammation of the
adjacent draining lymph vessels
 Up to 20% of all grown man in
certain communities in Haiti suffer
from hydrocele
 No effects on fertility, but wide
spread sexual disability
 Profound effects on patients self
esteem and family life

16. lymphatic filariasis
Adult filaria can be
detected in the scrotal
lymph vessels of men with
hydrocele by ultra sound
Macrofilaria can be
identified easily by there
mobility

17. lymphatic filariasis
 Chronic disease has complex
inflammatory etiology
 Dieing filaria are especially potent in
triggering inflammation, and death of
worms if followed by episodes of fever,
pain and acute inflammation
 Recent work suggests that, in addition to
adult worms, secondary bacterial and
fungal infections play an important role
in acute episodes and chronic
progression of the disease

18. lymphatic filariasis
Only a minority of
patients progresses to
elephantiasis and the
factors contributing to
their susceptibility are
not well understood
©WHO-TDR

19. lymphatic filariasis
Diagnosis:
 Demonstration of microfilaria in blood, or lymph (has to be done at
night!)
 Antibody and antigen capture assays (dip stick format)
 Demonstration of adult worms by ultra sound

20. lymphatic filariasis
Oedema of arm and hand in this patient with
filariasis
Lesions between fingers and toes are especially
vulnerable to bacterial infection

21. lymphatic filariasis
 Chemotherapy can be used to kill microfilaria but does not
affect adult worms (antibiotic treatment targeting Wolbachia
may hold promise here)
 However strict antiseptic regimens using soap and antibacterial
ointments can greatly reduce swelling, pain and other
symptoms and even revert many severe symptoms

22. sensit
Filariasis
Antibody
test

23. ubio’s solution for Filariasis
Diagnosis
One step Immuno-chromatographic test for Filarial
antibodies .
Whole blood or serum
Uses a novel recombinant filarial antigen.
Results in 10 minutes.
Filariasis can be detected in early stage of infection.

24. TEST PRINCIPLE
 The ubio sensit Filaria antibody Rapid Test is a lateral flow chromatographic
immunoassay.
 The test cassette consists of 1) a pink colored conjugate pad containing Anti Human IgG
conjugated with colloid gold, 2) a nitrocellulose membrane strip containing test band
(T) and a control band (C). The T band is precoated with recombinant Filaria Antigen for
the detection of anti-Filaria antibody.
 Test specimen, with adequate amount of buffer, migrates along the conjugate pad and
further across the coated membrane by capillary action.
 Anti-Filarial antibodies present in the sample, complex with the Anti Human IgG present
in the conjugate pad and gets captured onto the coated Filaria antigen. Thus giving a
colored test band, indicating a W. bancrofti or B. malayi positive test result.
 If the respective antibody is not present in the sample, no reaction occurs and no test line
is formed.
 The sample then migrates further along the strip until it reaches the control band, where
excess Anti Human IgG - colloidal gold gets bound and produces a second visible line on
the membrane. This control line indicates that the sample has migrated across the
membrane as intended

25. Test Procedure
Take out the test card from the foil pouch and place it on
a horizontal surface.
Add 10 μl of the specimen to the sample hole (marked
S) on the test card.
When the sample is fully absorbed, add three drops of
the diluents provided with the assay
Wait for 10-15 minutes and interpret results. The result
is considered invalid after 20 minutes. All results where
control band does not appear are considered invalid.

27. Test Performance
100 % specificity, (in endemic and non
endemic normal)
no cross reaction with Leishmania, malaria
and other nematode parasites.
>97% Sensitivity.

28. About ubio
Leading IVD manufacturer from India
ISO 13485 and GMP certified Manufacturing Facility.
Focus on Neglected Tropical Diseases.
Products include: Malaria pf, Malaria Pv, Malaria
pf/Pan, Malaria Pf/Pv, TB MPT 64, TB LAM,
Leishmania, Filaria, Leptospira IgM, Salmonella IgM,
Brucella IgG, H.Pylori Ab/Ag, hCG and LH tests
Veterinary Diagnostics.
Leading uncut sheet supplier.
OEM and Bulk manufacture.

29. For further details and placing your orders
ubio Biotechnology Systems Pvt Ltd.,
XII-111E/F, Biotechnology Incubation Center,
KINFRA Hi-tech Park, Kalamassery,
Cochin – 683503, Kerala, India
Ph: +91-484-2532966, +91-9744122269
www.ubio.in
contact@ubio.in