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Diagnosis of Human Filariasis
ubio sensit Filariasis Antibody Test
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
Lymphatic Filariasis: Life Cycle
Filaria and filariasisTissue dwelling nematodes with adults and
L1 present in the human hostArthropode vector which takes up L1 and
transmits L3Onchocerciasis, Lymphatic filariasis,
(Loiasis)
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
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
lymphatic filariasis
Wucheria bancrofti
Brugia malayi
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)
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
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)
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 midgut wall and migrate to the flight muscle where they molt twice (the develop intra-cellularly)
Culex quiquefasciatus
W. bancrofti L1 in mosquito flight muscle
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
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
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
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
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
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
lymphatic filariasis
Only a minority of patients progresses to elephantiasis and the factors contributing to their susceptibility are not well understood©WHO-TDR
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
lymphatic filariasis
Oedema of arm and hand in this patient with filariasis
Lesions between fingers and toes are especially vulnerable to bacterial infection
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
sensit FilariasisAntibody
test
ubio’s solution for Filariasis Diagnosis
One step Immuno-chromatographic test for Filarial antibodies .
Whole blood or serumUses a novel recombinant filarial
antigen.Results in 10 minutes.Filariasis can be detected in early stage
of infection.
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
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.
Test Performance100 % specificity, (in endemic
and non endemic normal)no cross reaction with
Leishmania, malaria and other nematode parasites.
>97% Sensitivity.
About ubio
Leading IVD manufacturer from IndiaISO 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.
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, IndiaPh: +91-484-2532966, +91-