Loop-Mediated Isothermal Amplification for Detection of the 5.8S Ribosomal Ribonucleic Acid Internal Transcribed Spacer 2 Gene Found inTrypanosoma brucei gambiense.
The loop-mediated isothermal amplification (LAMP) assay with its advantages of cost effectiveness, rapidity, and simplicity, has evolved as a sensitive and specific method for the detection of African trypanosomes. Highly sensitive LAMP reactions specific forTrypanosoma brucei rhodesienseor that recognize but do not discriminate betweenTrypanosoma brucei brucei,T. b. rhodesiense,Trypanosoma brucei gambiense, andTrypanosoma evansihave been developed. A sensitive LAMP assay targeting theT. b. gambiense5.8S ribosomal RNA internal transcribed spacer 2 (5.8S-ITS2) gene is also available but this assay does not target binding sites that span the CCCA (C3A) (557-560 bps) insertion site that further differentiatesT. b. gambiensefromT. b. bruceiHere we describe 5.8S-ITS2-targeted LAMP assay that fit these criteria. The LAMP primer sets containing theT. b. gambiense-specific C3A tetranucleotide at the start of the outer forward primer sequences showed high specificity and sensitivity down to at least 0.1 fgT. b. gambiensegenomic DNA.
Department of Pathology, The Johns Hopkins University School of Medicine, Baltimore, Maryland.
Olga V
OV
Nikolskaia
Unit for Environmental Sciences and Management, North-West University, Potchefstroom, South Africa.
Oriel M M
OM
Thekisoe
Department of Pathology, University of Maryland School of Medicine, Baltimore, Maryland.
J Stephen
JS
Dumler
Department of Pathology, The Johns Hopkins University School of Medicine, Baltimore, Maryland. dgrab1@jhmi.edu.
Dennis J
DJ
Grab
D042241:Early Diagnosis; D017125:Genes, Protozoan / Q000235:genetics*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D012340:RNA, Ribosomal, 5.8S / Q000235:genetics*; D014347:Trypanosoma brucei gambiense / Q000235:genetics*; D014353:Trypanosomiasis, African / Q000175:diagnosis* / Q000469:parasitology
D012340:RNA, Ribosomal, 5.8S