Evaluation of copromicroscopy, multiplex-qPCR and antibody serology for monitoring of human ascariasis in endemic settings

PLoS Negl Trop Dis. 2024 Jun 18;18(6):e0012279. doi: 10.1371/journal.pntd.0012279. eCollection 2024 Jun.

Abstract

Background: The standard diagnosis of Ascaris lumbricoides and other soil-transmitted helminth (STH) infections relies on the detection of worm eggs by copromicroscopy. However, this method is dependent on worm patency and shows only limited accuracy in low-intensity infection settings. We aimed to decipher the diagnostic accuracy of different antibodies using various Ascaris antigens in reference to copromicroscopy and quantitative PCR (qPCR), four months after national STH preventative chemotherapy among school children in western Kenya.

Methodology: STH infection status of 390 school children was evaluated via copromicroscopy (Kato-Katz and mini-FLOTAC) and qPCR. In parallel, Ascaris-specific antibody profiles against larval and adult worm lysates, and adult worm excretory-secretory (ES) products were determined by enzyme-linked immunosorbent assay. Antibody cross-reactivity was evaluated using the closely related zoonotic roundworm species Toxocara cati and Toxocara canis. The diagnostic accuracy of each antibody was evaluated using receiver operating curve analysis and the correspondent area under the curve (AUC).

Principal findings: Ascaris was the predominant helminth infection with an overall prevalence of 14.9% (58/390). The sensitivity of mini-FLOTAC and Kato-Katz for Ascaris diagnosis reached only 53.5% and 63.8%, respectively compared to qPCR. Although being more sensitive, qPCR values correlated with microscopic egg counts (R = -0.71, P<0.001), in contrast to antibody levels. Strikingly, IgG antibodies recognizing the ES products of adult Ascaris worms reliably diagnosed active Ascaris infection as determined by qPCR and microscopy, with IgG1 displaying the highest accuracy (AUC = 0.83, 95% CI: 0.75-0.91).

Conclusion: IgG1 antibody responses against adult Ascaris-ES products hold a promising potential for complementing the standard fecal and molecular techniques employed for monitoring Ascaris infections. This is of particular importance in the context of deworming programs as the antibody diagnostic accuracy was independent of egg counts.

Publication types

  • Evaluation Study

MeSH terms

  • Adolescent
  • Animals
  • Antibodies, Helminth* / blood
  • Antigens, Helminth / immunology
  • Ascariasis* / diagnosis
  • Ascariasis* / epidemiology
  • Ascariasis* / immunology
  • Ascaris / immunology
  • Ascaris / isolation & purification
  • Ascaris lumbricoides / immunology
  • Ascaris lumbricoides / isolation & purification
  • Child
  • Endemic Diseases
  • Enzyme-Linked Immunosorbent Assay / methods
  • Feces* / parasitology
  • Female
  • Humans
  • Kenya / epidemiology
  • Male
  • Microscopy / methods
  • Multiplex Polymerase Chain Reaction / methods
  • Real-Time Polymerase Chain Reaction / methods
  • Sensitivity and Specificity*

Substances

  • Antibodies, Helminth
  • Antigens, Helminth

Grants and funding

This work was supported by a grant from Einstein Foundation Berlin and the internationalization booster of Freie University Berlin to SH, Deutsche Forschungsgemeinschaft (DFG) Research Training Group 2046 to SH, FE and SR. RM received a monthly stipend from the funder “Einstein Foundation Berlin”. The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.