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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 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)
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
 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
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
Filariasis
Antibody
    test
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.
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 Performance
    100 % 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 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.
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

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ubio sensit Filariasis Antibody rapid test

  • 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
  • 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
  • 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
  • 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.
  • 26.
  • 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

Editor's Notes

  1. Numerous calcified nodes containing W. bancrofti mafi
  2. Numerous calcified nodes containing W. bancrofti mafi
  3. Numerous calcified nodes containing W. bancrofti mafi
  4. Numerous calcified nodes containing W. bancrofti mafi
  5. Numerous calcified nodes containing W. bancrofti mafi