Limits...
Development of chemiluminescent lateral flow assay for the detection of nucleic acids.

Wang Y, Fill C, Nugen SR - Biosensors (Basel) (2012)

Bottom Line: Rapid, sensitive detection methods are of utmost importance for the identification of pathogens related to health and safety.On-membrane enzymatic signal amplification is used to reduce the limit of detection to the sub-femtomol level.The limit of detection was determined to be 0.5 fmols of nucleic acid target.

View Article: PubMed Central - PubMed

Affiliation: Department of Food Science, University of Massachusetts, 102 Holdsworth Way, Amherst, MA 01003, USA. yuhongw@foodsci.umass.edu.

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.

No MeSH data available.


Related in: MedlinePlus

The CCD image of the lateral flow test strip tested in luminol-H2O2-HRP-chemiluminescent system taken by using a CCD imager. The image shows the low limit of detection with undetectable background signal using a luminol-based chemiluminescent system.
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biosensors-02-00032-f003: The CCD image of the lateral flow test strip tested in luminol-H2O2-HRP-chemiluminescent system taken by using a CCD imager. The image shows the low limit of detection with undetectable background signal using a luminol-based chemiluminescent system.

Mentions: The effect of the target concentration on the chemiluminescent signal was visualized with a CCD camera (Figure 3). It can be seen that while the control line was present in all target levels tested, the signal generated at the test line decreased noticeably as the target levels decreased. This response was expected as the decrease in target would result in a decrease in HRP on the test line.


Development of chemiluminescent lateral flow assay for the detection of nucleic acids.

Wang Y, Fill C, Nugen SR - Biosensors (Basel) (2012)

The CCD image of the lateral flow test strip tested in luminol-H2O2-HRP-chemiluminescent system taken by using a CCD imager. The image shows the low limit of detection with undetectable background signal using a luminol-based chemiluminescent system.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263543&req=5

biosensors-02-00032-f003: The CCD image of the lateral flow test strip tested in luminol-H2O2-HRP-chemiluminescent system taken by using a CCD imager. The image shows the low limit of detection with undetectable background signal using a luminol-based chemiluminescent system.
Mentions: The effect of the target concentration on the chemiluminescent signal was visualized with a CCD camera (Figure 3). It can be seen that while the control line was present in all target levels tested, the signal generated at the test line decreased noticeably as the target levels decreased. This response was expected as the decrease in target would result in a decrease in HRP on the test line.

Bottom Line: Rapid, sensitive detection methods are of utmost importance for the identification of pathogens related to health and safety.On-membrane enzymatic signal amplification is used to reduce the limit of detection to the sub-femtomol level.The limit of detection was determined to be 0.5 fmols of nucleic acid target.

View Article: PubMed Central - PubMed

Affiliation: Department of Food Science, University of Massachusetts, 102 Holdsworth Way, Amherst, MA 01003, USA. yuhongw@foodsci.umass.edu.

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.

No MeSH data available.


Related in: MedlinePlus