Utilization of Nucleotide Probes in PCR‐ELISA Procedure for the Quantitative Determination of Plasmodium Falciparum DNA in Malaria

Abstract

The research work reported herein is the development of a simple and specific quantitative procedure for the determination of P. falciparum DNA in malaria that involves the direct detection of the highly 42‐kDa conserved C‐terminal regiopn of P. falciparum merozoite surface protein gene (MSP1 ₄₂ gene). This procedure entails the amplification of the MSP1 ₄₂ 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 MSP1 ₄₂ gene. These specific probes are then used in the Enzyme Linked Immunosorbent Assay (ELISA) for the quantitative determination of the MSP1 ₄₂ gene which leads to the quantitative determination of P. falciparum DNA in malaria for quantitative diagnostic purpose. The P. falciparum malaria diagnostic results obtained from a small number of 18 whole blood samples show that the present quantitative PCR‐ELISA procedure allows the quantitative determination of P. falciparum DNA in malaria with a sensitivity and specificity over to those of the current standard microscopic examination. This quantitative PCR‐ELISA procedure is not only important for quantitative 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.

Keywords:

• Plasmodium falciparum
• MSP1 ₄₂ gene
• polymerase chain reaction
• enzyme linked immunosorbent assay
• biotin label nucleotide probes
• immobilized streptavidin polystyrene microtitration plates