The research work reported herein is the development of a simple and specific quant. procedure for the determination of P. falciparum DNA in malaria that involves the direct detection of the highly 42-kDa conserved C-terminal region of P. falciparum merozoite surface protein gene (MSP1₄₂ gene).This procedure entails the amplification of the MSP1₄₂ gene by using the PCR technique in the presence of digoxigenin-11-dUTP and the synthesis of the specific biotin label nucleotide probes directed to the MSP1₄₂ gene.These specific probes are then used in the Enzyme Linked Immunosorbent Assay (ELISA) for the quant. determination of the MSP1₄₂ gene which leads to the quant. determination of P. falciparum DNA in malaria for quant. diagnostic purpose.The P. falciparum malaria diagnostic results obtained from a small number of 18 whole blood samples show that the present quant. PCR-ELISA procedure allows the quant. determination of P. falciparum DNA in malaria with a sensitivity and specificity over to those of the current standard microscopic examinationThis quant. PCR-ELISA procedure is not only important for quant. P. falciparum malaria diagnosis but also useful for monitoring the efficacy of any existing anti-malarial drug as well as for testing the efficacy of any malaria vaccine.

01 Jan 2006·Analytical Letters

Selection of Peptide Ligands Specific for Baculovirus DNA‐Binding Protein from the Flitrx™ Random Peptide Display Library

Author: Nguyen, Khue Vu

Autographa californica nucleopolyhedrovirus (AcNPV) is a baculovirus that is widely employed as a vector for the expression of foreign genes and pest control.Although baculoviruses, including AcNPV, efficiently replicate in the nuclei of arthropod cells, the dynamics and mechanism of DNA replication within the infected cell are still poorly understood.It has been found that the DNA-binding protein (DBP) is an early gene product and appears to be crucial for viral DNA replication.Presented here is the selection of peptide ligands that specifically bind to DBP for AcNPV from the FliTrx random peptide display library; this entails the amplification, cloning of the DNA-binding protein (DBP) gene from AcNPV and the construction of the expression plasmid for DBP, and the expression and purification of the recombinant His.Tag AcNPV DBP that was used as a target mol. for the selection of the peptide ligands specific for AcNPV DBP.The affinity and efficiency of such peptide ligands were then measured by ELISA procedures.The beneficial aspect of this research is the monospecificity quality of the peptide ligands specific for AcNPV DBP.They could be used for the study of the dynamics of the viral genome and its replication within the infected cell, for the development of a quant. method for the determination of the presence of baculovirus in various samples, for the development of a peptide ligand based assay for the determination of baculovirus titers; or they could be immobilized on a chromatog. support for an improved affinity purification of AcNPV DBP.

Biotechnology Series

Practical matters in instrumentation for mammalian cell cultures

Author: Fleischaker, Robert J. Jr.

A review with 8 references on some of the instrumental methods currently used in cell culture and the implementation and limitation of process control technol. as applied to these systems.

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