Leishmania Promastigote Membrane Antigen-Based Enzyme-Linked Immunosorbent Assay and Immunoblotting for Differential Diagnosis of Indian Post-Kala-Azar Dermal Leishmaniasis
IR@IICB: CSIR-Indian Institute of Chemical Biology, Kolkata
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| Title |
Leishmania Promastigote Membrane Antigen-Based Enzyme-Linked
Immunosorbent Assay and Immunoblotting for Differential
Diagnosis of Indian Post-Kala-Azar Dermal Leishmaniasis
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| Creator |
Saha, Samiran
Majumder, Tuhina Anam, Khairul Ravindran, Rajesh Bairagi, Bibhas Saha, Bibhuti Goswami, Ramapada Pramanik, Netai Guha, Subhasis K Kar, Sujata Banerjee, Diwadas Ali, Nahid |
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| Subject |
Infectious Diseases and Immunology
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| Description |
Diagnosis of post-kala-azar dermal leishmaniasis (PKDL), caused by Leishmania donovani, is difficult, as the
dermal lesions are of several types and resemble those caused by other skin diseases, especially leprosy. Since
the disease generally appears very late after the clinical cure of kala-azar in India, it is also difficult to correlate PKDL with a previous exposure to L. donovani. Very few attempts have been made so far to diagnose PKDL
serologically, and the diagnostic methods vary in their sensitivities and specificities. Diagnosis of PKDL
through sophisticated PCR methods, although highly sensitive, has limited practical use. We have developed
a serodiagnostic method using an enzyme-linked immunosorbent assay to detect specific immunoglobulin (Ig)
isotypes and IgG subclass antibodies in the sera of Indian PKDL patients. Our assay, which uses L. donovani
promastigote membrane antigens, was 100% sensitive for the detection of IgG and 96.7% specific for the
detection of IgG and IgG1. Optical density values for individual patients, however, demonstrated wide variations.
Western blot analysis based on IgG reactivity could differentiate patients with PKDL from control
subjects, which included patients with leprosy, patients from areas where kala-azar is endemic, and healthy
subjects, by the detection of polypeptides of 67, 72, and 120 kDa. The recognition patterns of the majority of
serum samples from patients with PKDL were also distinct from those of the serum samples from patients with
visceral leishmaniasis (VL), at least for a 31-kDa polypeptide. To further differentiate patients with PKDL
from those with active and cured VL, we analyzed the specific titers of the Ig isotypes and IgG subclasses. High
levels of IgG, IgG1, IgG2, and IgG3 antibodies significantly differentiated patients with PKDL from patients cured of VL. The absence of antileishmanial IgE and IgG4 in patients with PKDL differentiated these patients
from those with active VL. These results imply intrinsic differences in the antibodies generated in the sera from
patients with PKDL and VL.
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| Date |
2005
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| Type |
Article
PeerReviewed |
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| Format |
application/pdf
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| Identifier |
http://www.eprints.iicb.res.in/1137/1/JOURNAL_OF_CLINICAL_MICROBIOLOGY%2C43_%2C_3%2C__1269%2D1277__[90].pdf
Saha, Samiran and Majumder, Tuhina and Anam, Khairul and Ravindran, Rajesh and Bairagi, Bibhas and Saha, Bibhuti and Goswami, Ramapada and Pramanik, Netai and Guha, Subhasis K and Kar, Sujata and Banerjee, Diwadas and Ali, Nahid (2005) Leishmania Promastigote Membrane Antigen-Based Enzyme-Linked Immunosorbent Assay and Immunoblotting for Differential Diagnosis of Indian Post-Kala-Azar Dermal Leishmaniasis. Journal of Clinical Microbiology, 43 (3). pp. 1269-1277. |
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| Relation |
http://dx.doi.org/10.1128/JCM.43.3.1269–1277.2005
http://www.eprints.iicb.res.in/1137/ |
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