Develop quantitative NASBA assay to detect E. coli.The nucleic acid nucleic acid, any of a group of organic substances found in the chromosomes of living cells and viruses that play a central role in the storage and replication of hereditary information and in the expression of this information through protein synthesis. sequence-based amplification (NASBA NASBA National Association of State Boards of Accountancy
NASBA Nucleic Acid Sequence-Based Amplification (assay used to detect HIV viral load in blood plasma) ) is a very sensitive and rapid technique for amplifying RNA RNA: see nucleic acid.
in full ribonucleic acid
One of the two main types of nucleic acid (the other being DNA), which functions in cellular protein synthesis in all living cells and replaces DNA as the carrier of genetic and for detecting bacteria. NASBA is a specific, isothermal i·so·ther·mal
Of, relating to, or indicating equal or constant temperatures.
having the same temperature. method of nucleic acid amplification which is highly suited for amplifying RNA. However, when using the NASBA, it is not possible to quantify the production yield of the tested target sequence.
Competitive assays have been developed to quantify the target sample. These assays use internal standards that compete with the target sequence for multiplication during the amplification reaction. One current drawback of these assays entails the considerable effort that is required to construct an appropriate RNA or DNA DNA: see nucleic acid.
or deoxyribonucleic acid
One of two types of nucleic acid (the other is RNA); a complex organic compound found in all living cells and many viruses. It is the chemical substance of genes. internal standard sequence using genetic engineering. Much effort has to be put into designing a sequence that can equally compete with the target sequence for amplification in the same reaction tube.
So, the development of a simple procedure that produces a competitor molecule is of great interest. Instead of cloning sequences, carrying out deletion-PCR reactions and sequencing the successful construct, Cornell University scientists were able to show that the careful design of NASBA-deletion primers can lead to a competitor molecule within only two NASBA reactions.
A quantitative competitive NASBA assay that determines the initial concentration of E. coli RNA can be achieved within 3 hours with the use of an electrochemiluminesence (ECL (Emitter-Coupled Logic) A digital circuit composed of bipolar transistors in which the emitter ends are wired together. ECL gates switch faster than TTL gates, but consume more power. See TTL, I2L and bipolar.
1. ) detection method and a simple lateral-flow assay. The ECL technique is extremely sensitive. Its dynamic range spans over several orders of magnitude.
Cornell researchers integrated a lateral-flow biosensor A device that detects and analyzes body movement, temperature or fluids and turns it into an electronic signal. See lab on a chip and data glove.
Biosensor into the internal standards onto the same assay. This made it possible to detect and quantify the target RNA in just one simple assay. Internal standards constructed by this method competed equally with the RNA of the target E. coli.
Further information. Antje Baeumner, Department of Biological and Environmental Engineering, Cornell University, 318 Riley-Robb Hall, Ithaca, NY 14853; phone: 607-255-5433; fax: 607-255-4080; email: firstname.lastname@example.org.