Deep kinetoplast genome analyses result in a novel molecular assay for detecting Trypanosoma brucei gambiense-specific minicircles

Manon Geerts, Zihao Chen, Nicolas Bebronne, Nicholas J. Savill, Achim Schnaufer, Philippe Buscher, Nick Van Reet, Frederik Van den Broeck

Research output: Contribution to journalA1: Web of Science-articlepeer-review

16 Downloads (Pure)

Abstract

The World Health Organization targeted Trypanosoma brucei gambiense (Tbg) human African trypanosomiasis for elimination of transmission by 2030. Sensitive molecular markers that specifically detect Tbg type 1 (Tbg1) parasites will be important tools to assist in reaching this goal. We aim at improving molecular diagnosis of Tbg1 infections by targeting the abundant mitochondrial minicircles within the kinetoplast of these parasites. Using Next-Generation Sequencing of total cellular DNA extracts, we assembled and annotated the kinetoplast genome and investigated minicircle sequence diversity in 38 animal- and human-infective trypanosome strains. Computational analyses recognized a total of 241 Minicircle Sequence Classes as Tbg1-specific, of which three were shared by the 18 studied Tbg1 strains. We developed a minicircle-based assay that is applicable on animals and as specific as the TgsGP-based assay, the current golden standard for molecular detection of Tbg1. The median copy number of the targeted minicircle was equal to eight, suggesting our minicircle-based assay may be used for the sensitive detection of Tbg1 parasites. Annotation of the targeted minicircle sequence indicated that it encodes genes essential for the survival of the parasite and will thus likely be preserved in natural Tbg1 populations, the latter ensuring the reliability of our novel diagnostic assay.

Original languageEnglish
Article number081
JournalNAR Genomics and Bioinformatics
Volume4
Issue number4
Pages (from-to)lqac081
Number of pages14
DOIs
Publication statusPublished - 2022

Keywords

  • SLEEPING SICKNESS
  • CONSERVED SEQUENCE
  • DNA MINICIRCLES
  • ORGANIZATION
  • IDENTIFICATION
  • PCR
  • CLONALITY
  • DIVERSITY
  • EVOLUTION
  • PARENTS

Fingerprint

Dive into the research topics of 'Deep kinetoplast genome analyses result in a novel molecular assay for detecting Trypanosoma brucei gambiense-specific minicircles'. Together they form a unique fingerprint.

Cite this