Comparison of a nested polymerase chain reaction – restriction fragment length polymorphism method, the PATH antigen detection method, and microscopy for the detection and identification of malaria parasites

Abstract

A nested polymerase chain reaction – restriction fragment length polymorphism (PCR–RFLP) method, the PATH antigen detection method, and light microscopy were compared for their capacity to detect and identify Plasmodium species. One hundred and thirty-six blood specimens obtained from patients suspected of having malaria were examined by each of the three methods. Forty-four specimens were positive for malaria using microscopy as the "gold standard". The sensitivity for nested PCR was 100%, and the specificity was 98%. For the detection of Plasmodium falciparum, the antigen detection method had a sensitivity of 100% and a specificity of 97%. Species identification obtained using PCR–RFLP was identical or superior to light microscopy in 42 cases (96%). Although the nested PCR–RFLP method was more sensitive and specific, the rapid turnaround time and high sensitivity of the antigen detection method makes it a useful adjunct to standard microscopy.Key words: malaria, PCR–RFLP, antigen detection.