Assay nucleic acid amplification

Antibody capture of viruses can be used as a preparatory step in nucleic acid amplification techniques. Immunocapture of virus particles can be used to streamline and/or optimize the concentration, purification and specificity requirements of polymerase chain reaction assays. 

All of these tasks can now be addressed with a family of novel tests collectively named product-enhanced reverse transcriptase (PERT) assays. PERT assays combine the broad detection range of RT tests with the high sensitivity of nucleic acid amplification procedures. PERT assays are based on the selective enhancement, by polymerase chain reaction (PCR) or one of the various... 

Baker CA, Cartwright CP, Williams DN> Nelson SM, Peterson PK. Early detection of central nervous system tuberculosis with the gen-probe nucleic acid amplification assay utility in an inner city hospital. Clin Infect Dis 2002 35 339-42. 

Saldanha J, Lelie N, Heath A. Establishment of the first international standard for nucleic acid amplification technology (NAT) assays for HCV RNA. WHO Collaborative Study Group. Vox Sang 1999 76 149-58. 

Strain specific lysis by bacteriophages can be used to release intracellular ATP or adenylate kinase (AK). At the 13 ISBC a paper is presented on identification of bacteria using bacteriophages. There are also several papers on bioluminescent realtime detection of nucleic acid amplification, which may be used for identification of bacteria. In these assays either pyrophosphate or AMP is converted to ATP as a measure of the amplification reaction. 

Recombinant luciferases and stable liquid ATP standards have increased the reliability of ATP biomass assays. Detection limits of ATP and ATP+AMP assays have reached the level of a single cell and can, combined with filtration techniques, reach 1 cell/mL. The assay of ATP+AMP is the most reliable method for hygiene monitoring. With immunocapture, bacteriophages or nucleic acid amplification combined with sensitive ATP reagents bacteria can be identified. In the near future we expect to see an abundance of user-friendly systems at acceptable prices. 

Novel Bioluminescent Assay for Real-Time Nucleic Acid Amplification... 

The first step in realising this concept required the development of an assay for the detection of 5 -AMP via firefly luciferase that can work solutions containing dNTPs and other components of a nucleic acid amplification reaction. A number of methods to detect NMPs via coupling to luciferase are known. Taking into account... 

Gandelman O, Church V, Murray JAH, Tisi LC. A single-step bioluminescent endpoint assay for nucleic acid amplification. This volume... 

Sutherland, J. W. Patterson, D. R. Homogeneous method for assay of double-stranded nucleic acid products in nucleic acid amplification using fluorescent dyes. Eur. Pat. Appl. EP 684316, 1995 Chem. Abstr. 1996, 124, 137781. 

Staining Applications Nucleic acids cells bacteria ° chromatin genes leukocytes nu-clei micronuclei megakaryocyte microorganisms peptides proteins antibodies sperms Biological Applications Nucleic acid hybridization nucleic acid amplification assay DNA fragment siz-... 

Morrison, T. Improved selective ligation and nucleic acid amplification assay. PCX Int. Appl. WO 2005059178, 2005 Chem. Abstr. 2005, 143, 72747. 

The molecular sensitivities of the first and second generations of the bDNA assays were limited by nonspecific hybridization between the amplification probes and other nucleic acids. Short regions of hybridization between any of the probes constituting the amplification system, (preamplifier, amplifier, and labeled probe) and any nontarget nucleic acid sequence leads to amplification of the background signal. Capture probes, capture extenders, and sample nucleic acid are all sources of this background hybridization (Collins et al 1997). 

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