DNA Detection and Quantification

Detection of the M. tuberculosis, HBV-DNA and HCV-RNA from the real samples by PCR 39 sputum collected from patients with suspected pulmonary tuberculosis, 157 sera collected from patients with HBV infection, and 16 sera collected from patients with HCV infection were selected in this study. The sputa forM tuberculosis detection and the sera for HBV-DNA detection and quantification were treated by BOOM reagents and by carbon nano-tube for DNA extraction. The sera for HCV-RNA detection and quantification were treated by Trizol-LS reagents and by carbon nano-tube for RNA extraction. The PCR was carried out for M. tuberculosis detection. The PCR and the qPCR were carried out for detection and quantification of HBV-DNA and HCV-RNA. The obtained results demonstrated that the BOOM method, Trizol-LS versus carbon nanotubes have given no difference 6/39 sputum were positive withM. tuberculosis, 103/157 sera were positive with HBV-DNA, 15/16 were positive with HCV-RNA. In qPCR, no difference more than 1 log was detected between carbon nano-tube method versus BOOM method and Trizol-LS... 

D. Redecker, T. Batinic, I. S. Feder, K. Koseh, U. Schulz, P. Vinuesa, and D. Werner, Biocontrol strain Pseudomomis fluorescens W34 specific detection and quantification in the rhizo.sphere of Cucumis salivus with a DNA probe and characterization by DNA fingerprinting. 2. Natmforsch. 54c 359 (1999). 

Pawlotsky, J.-M., et al. (1997). What technique should be used for routine detection and quantification of HBV DNA in clinical samples J. Virol. Methods 65,245-253. 

Computer-Based Methods for Detection and Quantification of Concerted Evolution in Ribosomal DNA Sequences... 

Once incorporated, unbound lewisite is quickly hydrolyzed. Its predominant metabolite is 2-chlorovinylarsonous acid, CVAA (Figure 50.8). Analytical methods to confinn lewisite exposure have, at least in the past, focused on the detection and quantification of CVAA. However, Noort et al. (2002) also pointed out that due to the high affinity of arsenic towards sulfhydryl groups, adducts of lewisite/ CVAA and cysteine residues of proteins are formed. In an in vitro study, incubating " C-labeled lewisite with human blood samples, 90% of lewisite was found in erythrocytes, whereas 25 to 50% of arsenic was bound to globin. From these protein adducts, CVAA can be released to form an adduct with the antidote British Anti-Lewisite (BAL) (Fidder et al, 2000). The authors were also able to identify a specific protein adduct of lewisite formed with the cysteine residues 93 and 112 of P-globin. See Detection of DNA and protein adducts of vesicants, below, for analytical... 

Frank, A. J., Proctor, S. J., and Tilby, M. J. (1996) Detection and quantification of melphalan-DNA adducts at the single cell level in hemopoietic tumor cells. Blood 88(3), 977-984. 

Caliendo AM, Yen-Lieberman B, Baptista J, Andersen J, Crumpacker C, Schuurman R, et al. Comparison of molecular tests for detection and quantification of cell-associated cytomegalovirus DNA. J Clin Microbiol 2003 41 3509-13. 

High-performance liquid chromatography (HPLC) separation of DNA adducts, followed by fluorescence or electrochemical detection and quantification. 

Muller, R., Adamkiewicz, J, and Rajewsky, M. F. (1982). Immunological detection and quantification of carcinogen-modified DNA components. lARC Sci Publ, 463 79. 

RNA detection and quantification with BART. Purified RNA from classical swine fever virus (obtained from FLI, Germany) was amplified in a one-step format which included reverse-transcription with AMV and LAMP. As with any DNA target for a wide dilution series of RNA samples BART resulted in a sequence of light peaks... 

However, the key technological bottleneck remains the detection and quantification of the amplified DNAs. 

Aptamers have a strong potential for multiplex sensing of proteins. Thus, a chip-based biosensor was developed for specific detection and quantification of cancer-associated proteins in complex biological mixtures using immobilized fluorescently labeled DNA and RNA aptamers. Fluorescence polarization anisotropy was used for solid- and solution-phase measurements of target protein binding [51]. 

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