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. 2022 Oct 3:117:e220086.
doi: 10.1590/0074-02760220086. eCollection 2022.

Immunoanalysis of different antigenic preparations of Angiostrongylus cantonensis for neuroangiostrongyliasis diagnosis improvement

Leyva Cecília Vieira de Melo  1   2 , Felipe Corrêa Rezende de Souza  1   2 , Amanda de Oliveira Baccin  1   2 , Dan Jessé Gonçalves da Mota  3 , Vera Lucia Pereira-Chioccola  2   4 , Pedro Luiz Silva Pinto  1   2
Affiliations

Immunoanalysis of different antigenic preparations of Angiostrongylus cantonensis for neuroangiostrongyliasis diagnosis improvement

Leyva Cecília Vieira de Melo et al. Mem Inst Oswaldo Cruz. .

Abstract

Background: Angiostrongylus cantonensis is the etiological agent of neuroangiostrongyliasis in humans, which is developed in gastropods and vertebrate species, mainly rodents. Human transmission occurs through consumption of molluscs and paratenic hosts infected with L3, and the migration of larvae to the central nervous system causes eosinophilic meningitis. Laboratory diagnosis is based on molecular and immunological tests, using young or adult females as a source of antigens. However, these tests give positive results only after several weeks of symptoms onset and also cross-reactions with others parasite infections may occur.

Objectives: The purpose of this work was to study different antigenic preparations of distinct evolutionary phases of A. cantonensis, in order to improve serological techniques for disease immunodiagnosis.

Methods: For this purpose, antigenic fractions of different evolutionary forms were evaluated by Dot-enzyme-linked immunosorbent assay (Dot-ELISA) and Western blot using serum samples.

Findings: All analysed fractions showed reactivity with serum samples from patients with neuroangiostrongyliasis, especially female membrane alkaline (FAM) and female soluble alkaline (FAS) fractions together with female soluble saline (FSS), improving the technique specificity.

Main conclusions: The results point to the possibility of use of raw female antigens in association with alkaline membrane antigens extracted from adult worms to aid in diagnosis and helps initiate neuroangiostrongyliasis surveillance and control actions.

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Figures

Fig. 1:
Fig. 1:. purification of Angiostrongylus cantonensis L1 larvae with collagen. A: emulsification of collagen with the 1st Rugai sediment; B: 2nd Rugai in cold dechlorinated water; C: comparison between the sediments of the 1st (left) and 2nd Rugai (right); D: purified L1 larvae.
Fig. 2:
Fig. 2:. extraction of the reproductive system of adult females of Angiostrongylus cantonensis. A: identification of the sites for the sections (PE: excretory pore; TO: ovary terminations; V: vulva; An: anus); B: insertion of the needle into the anterior part of the worm; C: female after viscera extraction (CT = cuticle, and CV = visceral content); D: extracted female reproductive system.
Fig. 3:
Fig. 3:. scheme of protein fractions of extracts obtained from Angiostrongylus cantonensis. FAS: female soluble alkaline; MAS: male soluble alkaline; L1AS: L1 soluble alkaline; FAM: female membrane alkaline; MAM: male membrane alkaline; L1AM: L1 membrane alkaline; FSS: female soluble saline; MSS: male soluble saline; L1SS: L1 soluble saline; FSM: female membrane saline; MSM: male membrane saline; L1SM: L1 membrane saline; L3SM: L3 membrane saline; UTSS: soluble saline of uterus and ovaries; UTSM: Saline membrane of uterus and ovaries.
Fig. 4:
Fig. 4:. examples of Dot-enzyme-linked immunosorbent assay (ELISA) results on reactive (R) and non-reactive (NR) samples.
Fig. 5:
Fig. 5:. scheme proposed for a screening test using different Angiostrongylus cantonensis female antigens, using Dot-enzyme-linked immunosorbent assay (ELISA) technique, regarding the possibility that the patient presents neuroangiostrongyliasis.
See this image and copyright information in PMC

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