Assessment of pan-Leishmania detection by recombinase polymerase amplification assay

Louizi C 1 , Khan MAA 2 , Faisal K 3 , Chowdhury R 3 , Ghosh P 3 , Hossain F 3 Show all authors , Nisansala T 4 , Ranasinghe S 5 , Moreno J 6 , Alvar J 7 , Mondal D 8 , Buhl T 9 , Lüder CGK 1 , Abd El Wahed A 10

Affiliations 

  • 1 Institute for Medical Microbiology and Virology, University Medical Center Goettingen, Georg-August University, Göttingen, Germany
  • 2 Institute of Animal Hygiene and Veterinary Public Health, University of Leipzig, Leipzig, Germany; Nutrition and Clinical Services Division, International Centre for Diarrheal Disease Research Bangladesh, Dhaka, Bangladesh. Electronic address: [email protected]
  • 3 Nutrition and Clinical Services Division, International Centre for Diarrheal Disease Research Bangladesh, Dhaka, Bangladesh
  • 4 Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, Pengkalan Chepa, Kota Baru, Kelantan, Malaysia; Faculty of Medical Sciences, University of Sri Jayewardenepura, Gangodawila, Nugegoda, Sri Lanka
  • 5 Faculty of Medical Sciences, University of Sri Jayewardenepura, Gangodawila, Nugegoda, Sri Lanka
  • 6 WHO Collaborating Center for Leishmaniasis, National Center for Microbiology, Instituto de Salud Carlos III, Madrid, Spain
  • 7 Royal Academy of Medicine of Spain, Madrid, Spain
  • 8 Nutrition and Clinical Services Division, International Centre for Diarrheal Disease Research Bangladesh, Dhaka, Bangladesh; Laboratory Sciences and Services Division, International Centre for Diarrheal Disease Research Bangladesh, Dhaka, Bangladesh
  • 9 Department of Dermatology, Venereology and Allergology, University Medical Centre Göttingen, Georg-August University, Göttingen, Germany
  • 10 Institute of Animal Hygiene and Veterinary Public Health, University of Leipzig, Leipzig, Germany
Diagn Microbiol Infect Dis, 2023 Feb;105(2):115862.
PMID: 36493571 DOI: 10.1016/j.diagmicrobio.2022.115862

Abstract

The spread of vector habitats along with increasing human mobility can introduce atypical Leishmania species and hence can challenge existing diagnostic practices for rapid detection of active infection with species outside the narrow target range. Here we assessed the pan-Leishmania detection ability of isothermal recombinase polymerase amplification (RPA) assays targeting 18S rRNA gene, cathepsin L-like cysteine proteinase B (Cpb) gene, and kinetoplast minicircle DNA (kDNA) regions. While the lowest limit of detection of the 18S rRNA-RPA and Cpb-RPA assays were estimated as 12 and 17 standard DNA molecules, respectively, both assays could amplify genomic DNA of 7 pathogenic Leishmania species. Evaluation of 18S rRNA-RPA and our previously developed kDNA-RPA assays on 70 real-time PCR-positive leishmaniasis samples of varying pathologies resulted in sensitivity rates of 35.71% and 88.57%, respectively, while the combined sensitivity was 98.57%. Combinatorial application of 18S rRNA-RPA and kDNA-RPA assays can be recommended for further diagnostic assessments.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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