UTILIZATION OF NUCLEOTIDE PROBES IN ELISA PROCEDURE FOR THE QUANTITATIVE DETERMINATION OF PLASMODIUM FALCIPARUM DNA IN MALARIA

Abstract

The present invention is the development of a simple and specific quantitative method 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 (MSP1) gene. This procedure entails the amplification of the 42-kDa C-terminal region of the MSP1 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 42-kDa C-terminal region of the MSP1 gene. These specific probes are then used in the Enzyme Linked Immunosorbent Assay (ELISA) for the quantitative determination of the 42-kDa C-terminal region of the MSP1 gene which leads to the quantitative determination of P. falciparum DNA in malaria for quantitative diagnostic purpose as well as for monitoring the efficacy of antimalarial treatment.

Description

Claims

1. The procedure using the specific biotin label nucleotide probes directed to the highly 42-kDa conserved C-terminal region of Plasmodium falciparum merozoite surface protein1 (MSP1) gene as a target for the development of a simple, specific and easy quantitative method for the determination of Plasmodium falciparum DNA in malaria, comprising: The procedure for amplifying the 42-kDa C-terminal region of the MSP1 gene from the negative control (non-infected) and P. falciparum infected samples, based on the polymerase chain reaction (PCR) in the presence of digoxigenin-11-dUTP using the synthesized oligonucleotides (SEQ ID NO. 1) and (SEQ ID NO. 2) for PCR:Isolating of P. falciparum DNA from the negative control and P. falciparum infected samples;Using the oligonucleotides