Evaluation of LAMP for the diagnosis of Loa loa infection in dried blood spots compared to PCR-based assays and microscopy

Mem Inst Oswaldo Cruz. 2022 Feb 11:116:e210210. doi: 10.1590/0074-02760210210. eCollection 2022.

Abstract

Background: Loa loa is a filarial species found exclusively in West and Central Africa. Microscopy is the traditional diagnosis method for human loiasis. Several molecular methods have developed as an alternative approach for identification of L. loa filarial parasites.

Objectives: The aim of this study was to evaluate a Loa-Loop-mediated isothermal amplification (LAMP) assay to diagnose loiasis disease on dried blood spots (DBS) samples, compared to microscopy, filaria-real time-polymerase chain reaction (PCR) and nested-Loa PCR.

Methods: A total of 100 DBS samples and 100 blood smears were used for this study. DNA was extracted using saponin/Chelex method. DNA isolated was assayed by a Loa-LAMP assay in parallel to microscopy, filaria-real time PCR and nested-Loa PCR. The sensitivities and specificities of Loa-LAMP assay was computed comparing to each one of the reference methods.

Findings: Loa-LAMP's sensitivity was more than 90% and specificity was nearly 100% when compared to molecular methods. On the other hand, sensitivity was decreased a bit when Loa-LAMP faced microscopy, but keeping the other statistical values high.

Main conclusions: Loa-LAMP is an appropriate method for loiasis diagnosis in endemic areas. Though, it has disadvantages like the reagents' high price at the moment and not to be able to detect more filarial species at once.

MeSH terms

  • Humans
  • Loiasis* / diagnosis
  • Microscopy
  • Molecular Diagnostic Techniques
  • Nucleic Acid Amplification Techniques
  • Real-Time Polymerase Chain Reaction
  • Sensitivity and Specificity

Supplementary concepts

  • LAMP assay

Grants and funding

Fundación Estatal, Salud, Infancia y Bienestar Social (Institute of Health Carlos III) and Cooperative Research Network on Tropical Diseases. T-HT-T is hired at the Malaria and Neglected Tropical Diseases Laboratory of National Centre of Tropical Medicine with a contract named ISCIII-SARA BORRELL CD17CIII/00018, financed by the Institute of Health Carlos III