A single test approach for accurate and sensitive detection and taxonomic characterization of Trypanosomes by comprehensive analysis of internal transcribed spacer 1 amplicons

PLoS Neglected Tropical Diseases
Alex Kiarie GaithumaChihiro Sugimoto

Abstract

To improve our knowledge on the epidemiological status of African trypanosomiasis, better tools are required to monitor Trypanosome genotypes circulating in both mammalian hosts and tsetse fly vectors. This is important in determining the diversity of Trypanosomes and understanding how environmental factors and control efforts affect Trypanosome evolution. We present a single test approach for molecular detection of different Trypanosome species and subspecies using newly designed primers to amplify the Internal Transcribed Spacer 1 region of ribosomal RNA genes, coupled to Illumina sequencing of the amplicons. The protocol is based on Illumina's widely used 16s bacterial metagenomic analysis procedure that makes use of multiplex PCR and dual indexing. Results from analysis of wild tsetse flies collected from Zambia and Zimbabwe show that conventional methods for Trypanosome species detection based on band size comparisons on gels is not always able to accurately distinguish between T. vivax and T. godfreyi. Additionally, this approach shows increased sensitivity in the detection of Trypanosomes at species level with the exception of the Trypanozoon subgenus. We identified subspecies of T. congolense, T. simiae, T. vivax, and T...Continue Reading

References

Nov 1, 1992·International Journal for Parasitology·D K MasigaW C Gibson
Oct 1, 1990·Acta Tropica·I MaudlinP Milligan
Feb 1, 1981·Molecular and Biochemical Parasitology·C CastroM Castañeda
Mar 17, 2004·Veterinary Parasitology·J M NgairaN K Olembo
Dec 25, 2004·Parasitology Research·Z K NjiruA M R Dávila
Aug 2, 2005·Experimental Parasitology·Andrew CoxSusan Welburn
Sep 2, 2005·Tropical Medicine & International Health : TM & IH·P Van den BosscheR De Deken
Sep 15, 2005·Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases·Z K NjiruW C Gibson
Apr 25, 2007·International Journal for Parasitology·W Gibson
Jul 24, 2007·Experimental Parasitology·Kim PicozziSusan C Welburn
Sep 11, 2007·Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases·E R AdamsW C Gibson
Oct 30, 2007·Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases·P B HamiltonW C Gibson
Feb 5, 2008·Proceedings of the National Academy of Sciences of the United States of America·De-Hua LaiJulius Lukes
Feb 29, 2008·PLoS Medicine·Pere P SimarroPierre Cattand
Dec 17, 2009·BMC Bioinformatics·Christiam CamachoThomas L Madden
Mar 23, 2010·Trends in Parasitology·Peter Van den BosscheJérémy Bouyer
Apr 13, 2010·Trends in Parasitology·Emily R AdamsWendy C Gibson
Apr 8, 2011·Nucleic Acids Research·Ivica Letunic, Peer Bork
Jun 9, 2012·Parasites & Vectors·Lori PeacockWendy Gibson
Oct 25, 2012·PLoS Neglected Tropical Diseases·Harriet AutySusan C Welburn
Nov 13, 2012·PLoS Neglected Tropical Diseases·Pere P SimarroJean G Jannin
Jan 19, 2013·Molecular Biology and Evolution·Kazutaka Katoh, Daron M Standley
Apr 14, 2016·Parasites & Vectors·Pascal GrébautGustave Simo
May 26, 2017·Genome Biology and Evolution·Bart CuypersStijn Deborggraeve
Jul 10, 2017·Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases·Herakles A GarciaMarta M G Teixeira

❮ Previous
Next ❯

Citations

Mar 21, 2020·Scientific Reports·Alex GaithumaChihiro Sugimoto
Jan 16, 2021·Parasites & Vectors·William ShereniGiuliano Cecchi
Feb 16, 2021·Frontiers in Veterinary Science·Yukiko NakamuraJunya Yamagishi
Jun 7, 2021·Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases·Faith I EbhodagheMichael N Okal
Jul 24, 2021·Journal of Parasitic Diseases : Official Organ of the Indian Society for Parasitology·Rotimi Johnson OjoMeshack Anthony Kugama
May 19, 2020·International Journal for Parasitology. Parasites and Wildlife·David SquarreJunya Yamagishi

❮ Previous
Next ❯

Datasets Mentioned

BETA
AB742530
KC196703
TVU22316
TCU22317
SRP159480
U22318

Methods Mentioned

BETA
PCR
PCRs
amplicon sequencing
dissection

Software Mentioned

LULU
Amptk
GraphPad
Primer
Simulate
Adobe Illustrator
UCLUST
BLAST
GraphPad Prism
MAFFT

Related Concepts

Related Feeds

African Trypanosomiasis

African trypanosomiasis, also known as sleeping sickness, is an insect-borne parasitic disease of humans and other animals. It is caused by protozoa of the species Trypanosoma brucei and almost invariably progresses to death unless treated. Discover the latest research on African trypanosomiasis here.