Performance of a Commercial Multiplex Allele-Specific Polymerase Chain Reaction Kit to Genotype African-Type Glucose-6-Phosphate Dehydrogenase Deficiency

Am J Trop Med Hyg. 2022 Dec 19;108(2):449-455. doi: 10.4269/ajtmh.21-1081. Print 2023 Feb 1.

Abstract

8-Aminoquinoline antimalarial drugs (primaquine, tafenoquine) are required for complete cure of Plasmodium vivax malaria, but they are contraindicated in patients with glucose-6-phosphate dehydrogenase (G6PD) deficiency. In the absence of spectrophotometry, which is a gold standard for measuring G6PD activity, G6PD genotyping is one of the alternatives to establish a database and distribution map of G6PD enzyme deficiency in Mauritania, which has become a new epicenter of P. vivax malaria in West Africa. The aim of our study was to assess the performance of multiplex allele-specific polymerase chain reaction (PCR) (African-type Diaplex C™ G6PD kit) against PCR-restriction fragment length polymorphism and sequencing. Of 146 mutations associated with G6PD A- genotypes in 177 blood samples from Mauritanian patients, all but two samples were identified correctly using multiplex allele-specific PCR (100% sensitivity and 99% specificity; "almost perfect agreement" between allele-specific PCR and PCR-restriction fragment length polymorphism/sequencing, with a kappa coefficient of 0.977). Despite a suboptimal PCR protocol for dried blood spots and the inability of the commercial assay to predict unequivocally the G6PD enzyme level in heterozygous females, the African-type Diaplex C™ G6PD genotyping kit seemed to be a valuable screening tool for male subjects and for research purposes in resource-limited countries where spectrophotometer and DNA sequencing are not available.

MeSH terms

  • Alleles
  • Antimalarials* / therapeutic use
  • Female
  • Genotype
  • Glucosephosphate Dehydrogenase / genetics
  • Glucosephosphate Dehydrogenase Deficiency* / diagnosis
  • Glucosephosphate Dehydrogenase Deficiency* / genetics
  • Humans
  • Malaria, Vivax* / drug therapy
  • Male
  • Multiplex Polymerase Chain Reaction
  • Primaquine / therapeutic use

Substances

  • Primaquine
  • Glucosephosphate Dehydrogenase
  • Antimalarials