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Indian J Dermatol Venereol Leprol. 2019 Aug 9. doi: 10.4103/ijdvl.IJDVL_485_18. [Epub ahead of print]

Evaluation of anti-gal enzyme-linked immunosorbent assay for the diagnosis of Indian post-kala-azar dermal leishmaniasis.

Author information

1
Department of Laboratory Medicine, School of Tropical Medicine, Kolkata, West Bengal, India.
2
Department of Tropical Medicine, School of Tropical Medicine, Kolkata, West Bengal, India.
3
Department of Dermatology, Bankura Sammilani Medical College, Government of West Bengal, Kenduadihi, Bankura, West Bengal, India.

Abstract

Background:

Elimination of kala azar from India is challenging as there are potential reservoirs of Leishmania donovani in patients with post-kala-azar dermal leishmaniasis (PKDL). The vast repertoire of carbohydrate moieties on L. donovani is known to elicit specific and strong humoral responses in patients with kala azar.

Aim:

The present study was undertaken to evaluate the diagnostic performances of anti-gal antibodies using enzyme-linked immunosorbent assay for successful serological diagnosis of PKDL in Indian patients and to differentiate cases of past cured visceral leishmaniasis infections.

Methods:

We developed Gal enzyme-linked immunosorbent assay to measure specific anti-gal IgG isotype in the sera of 71 Indian patients with PKDL. The diagnostic efficacy of the newly developed assay was evaluated for precision, sensitivity and accuracy.

Results:

Gal2 enzyme-linked immunosorbent assay revealed three-fold increased anti-gal titers in 71 patients with active PKDL compared to controls. Subclass enzyme-linked immunosorbent assay analysis further revealed enhanced IgG2 and IgG3 anti-gal titers in patients with PKDL compared to control subjects. The rank order for specificity and sensitivity for IgG subclasses was IgG3>IgG2>IgG4>IgG1. The area under the curve values of 0.98 and 0.99 were obtained for IgG and IgG3 Gal2 enzyme-linked immunosorbent assays respectively. Overall sensitivity and specificity were 95.7% (95% CI: 88.1-99.1) and 98.1% (95% confidence interval: 90.1-99.9), and 98.5% (95% CI: 92.4-99.9) and 98.1% (95% CI: 90.1-99.9), respectively. Intra-assay coefficient of variation was 1.5% and inter-assay coefficient of variation was 11.7%.

Limitations:

The Gal2 enzyme-linked immunosorbent assay needs to be further investigated in mass surveys.

Conclusion:

Taken together, anti-gal titers detected through Gal2 enzyme-linked immunosorbent assay can serve as an effective diagnostic tool in disease elimination setting and help in better case management in endemic districts.

KEYWORDS:

Asialofetuin; Leishmania donovani; PKDL; enzyme-linked immunosorbent assay; porcine stomach mucin

PMID:
31397399
DOI:
10.4103/ijdvl.IJDVL_485_18
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