Species-specific diagnosis of human malaria by the polymerase chain reaction

Lim PKC¹, Tan SK¹, Kita K², Kojima S², Watanabe Y², Mak JW¹ 

1. Division of Molecular Pathology, Institute for Medical Research, Jalan Pahang, 50588 Kuala. Lumpur, Malaysia

2. Department of Parasitology, Institute of Medical Science, The University of Tokyo, 4-6-1, Minato-ku, Tokyo 108, Japan

CITATION: Lim PKC, Tan SK, Kita K, Kojima S, Watanabe Y, Mak JW. Species-specific diagnosis of human malaria by the polymerase chain reaction. International Medical Research Journal. 1997 Jun;1(1):69–72. 

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ABSTRACT

The polymerase chain reaction (PCR) with species-specific primers derived from a number of Plasmodium genes has been shown to be highly specific and sensitive for malaria diagnosis. In this study, nucleotide sequences derived from the  mitochondrial cytochrome c oxidase III (CO III) genes of P. falciparum and P. vivax were used in the PCR to develop species-specific diagnostic assays. P. falciparum primers F1 and F2 only amplified P. falciparum template DNA while P vivax primers VI and V2 amplified both DNA templates of P. vivax and P. cynomolgi. The sensitivity of the PCR with F1 and F2 primers was 0.5 parasite per PCR reaction. PCR performed on DNA samples from 58 malaria patients and 30 uninfected individuals using both primer sets showed that both assays gave 100% sensitivity and 100% specificity. Three samples from patients with mixed infections of P falciparum and P. vivax were also positive in both the PCR assays with the two primer sets. The ease of the DNA preparation and PCR procedures used in this study makes this rest potentially useful for malaria diagnosis in the field, particularly where both P. falciparum and P. vivax occur together.

KEYWORDS: PCR, malaria diagnosis, mitochondrial cytochrome c oxidase III gene