Loop-Mediated isothermal amplification for detection of the 5.8s ribosomal ribonucleic acid internal transcribed spacer 2 gene found in trypanosoma brucei gambiense
Date
2017Journal
American Journal of Tropical Medicine and HygienePublisher
American Society of Tropical Medicine and HygieneType
Article
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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 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.Sponsors
This research was supported in part by grants from the National Institutes of Health (5R01AI082695 and 1R21AI079282).Keyword
loop-mediated isothermal amplificationLAMP
Genes, Protozoan--genetics
RNA, Ribosomal, 5.8S--genetics
Trypanosoma brucei gambiense--genetics
Trypanosomiasis, African--diagnosis
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https://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/10713/11143ae974a485f413a2113503eed53cd6c53
10.4269/ajtmh.15-0288
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