Loop-Mediated isothermal amplification for detection of the 5.8s ribosomal ribonucleic acid internal transcribed spacer 2 gene found in trypanosoma brucei gambiense
JournalAmerican Journal of Tropical Medicine and Hygiene
PublisherAmerican Society of Tropical Medicine and Hygiene
MetadataShow full item record
AbstractThe 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 for Trypanosoma brucei rhodesiense or that recognize but do not discriminate between Trypanosoma brucei brucei, T. b. rhodesiense, Trypanosoma brucei gambiense, and Trypanosoma evansi have been developed. A sensitive LAMP assay targeting the T. b. gambiense 5.8S ribosomal RNA internal transcribed spacer 2 (5.8SITS2) gene is also available but this assay does not target binding sites that span the CCCA (C3A) (557-560 bps) insertion site that further differentiates T. b. gambiense from T. b. brucei. Here we describe 5.8S-ITS2-targeted LAMP assay that fit these criteria. The LAMP primer sets containing the T. 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 fg T. b. gambiense genomic DNA.
SponsorsThis research was supported in part by grants from the National Institutes of Health (5R01AI082695 and 1R21AI079282).
Keywordloop-mediated isothermal amplification
RNA, Ribosomal, 5.8S--genetics
Trypanosoma brucei gambiense--genetics
Identifier to cite or link to this itemhttps://www.scopus.com/inward/record.uri?eid=2-s2.0-85014571151&doi=10.4269%2fajtmh.15-0288&partnerID=40&md5=1b6675f04ba379bf53c7a4711c9fc2f3; http://hdl.handle.net/10713/11143
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