Abstract
Rapid, sensitive detection methods are of utmost importance for the identification of pathogens related to health and safety. Herein we report the development of a nucleic acid sequence-based lateral flow assay which achieves a low limit of detection using chemiluminescence. On-membrane enzymatic signal amplification is used to reduce the limit of detection to the sub-femtomol level. To demonstrate this assay, we detected synthetic nucleic acid sequences representative of Trypanosoma mRNA, the causative agent for African sleeping sickness, with relevance in human and animal health in sub-Saharan Africa. The intensity of the chemiluminescent signal was evaluated by using a charge-coupled device as well as a microtiter plate reader. We demonstrated that our lateral flow chemiluminescent assay has a very low limit of detection and is easy to use. The limit of detection was determined to be 0.5 fmols of nucleic acid target.
Highlights
Trypanosoma such as Trypanosoma brucei and Trypanosoma cruzi, the causative agents for the potentially fatal African sleeping sickness and Chagas disease, respectively, have important influence on human health
The currently accepted methods for the Biosensors 2012, 2 detection of Trypanosoma such as microscopic examination and xenodiagnoses have poor sensitivity and are labor-intensive and time-consuming [1,2]. Immunological methods such as enzyme-linked immunosorbent assay, immunochromatographic dipstick test, radioimmunosorbent assay, and immunofluorescence antibody test are rapid and sensitive but not specific [3,4,5,6,7,8,9]. Molecular methods such as PCR and real-time nucleic acid sequence-based amplification are very specific but expensive and time consuming, combination of PCR and chemiluminescence southern blot has been used to improve the sensitivity of the detection of Trypanosoma [10,11,12,13,14,15,16,17]
When the volume is too high, the nitrocellulose membrane “floods”, causing the luminol/H2O2 mixture to move up the sides of the holder device by capillary action, preventing proper migration up the lateral flow test strip
Summary
Trypanosoma such as Trypanosoma brucei and Trypanosoma cruzi, the causative agents for the potentially fatal African sleeping sickness and Chagas disease, respectively, have important influence on human health. The currently accepted methods for the Biosensors 2012, 2 detection of Trypanosoma such as microscopic examination and xenodiagnoses have poor sensitivity and are labor-intensive and time-consuming [1,2] Immunological methods such as enzyme-linked immunosorbent assay, immunochromatographic dipstick test, radioimmunosorbent assay, and immunofluorescence antibody test are rapid and sensitive but not specific [3,4,5,6,7,8,9]. These techniques are not implemented in Trypanosomiasis control programs due to the high cost of the equipment [14,16,18]
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
