Genotyping of Plasmodium falciparum isolates by amplification of glutamate-rich protein gene using polymerase chain reaction]

To develop a genotyping method based on amplifying glutamate-rich protein (GLURP) gene for the diagnosis and identification of Plasmodium falciparum.Two pairs of primers specific for GLURP gene of P. falciparum were designed and synthesized. R2 polymorphic domain of GLURP gene was amplified by nested PCR, which was applied to genotyping of P. falciparum isolates obtained from patients attending the malaria clinic at the village of Borai, Thailand.Conspicuous polymorphism of GLURP alleles in natural populations of P. falciparum was found. 290 GLURP alleles were detected in 154 P. falciparum infections. Among the above-mentioned alleles, 12 different GLURP genotypes were distinguished according to different DNA sizes. Of them, the most frequently found allele was a variant of 770 bp, the least allele was that of 1,100 bp. More than 43% of the patients were found to be infected with mixed alleles. No apparent change for frequencies of the 12 different alleles was found in the 9-month longitudinal study.A genotyping method is developed for the research of strain taxonomy and pathogenesis of malaria parasites.