Detection of Plasmodium falciparum and Wuchereria bancrofti infected bloodsamples using multiplex PCR

A rapid and sensitive multiplex PCR has been developed for the diagnosis of multiple parasitic infection in human blood. Infection is detected by a si ngle multiplex PCR reaction containing two pairs of oligonucleotide primers whereby each primer is specific for each parasite species. These primer se ts amplified 400 and 450-bp fragments for Wuchereria bancrofti and 208-bp f ragment for Plasmodium falciparum. The PCR products derived from each paras ite species were visualized in ethidium bromide-stained agarose gels, there fore allowing the rapid identification of any, or all, of the two human par asites, ii present, in a single amplification reaction. This multiplex PCR was very sensitive with the ability to detect the presence of as little as 10 pg of parasite DNA. The primers used in this multiplex PCR also showed h ighly specific amplification of each respective parasite DNA without the pr esence of non-specific and non-target PCR products. This multiplex PCR syst em was used to analyse 36 human blood samples of Myanmar workers in the end emic area at Tak Province, Thailand. Two samples showed the multiple infect ion, 27 samples were either infected with W, bancrofti or P. falciparum and seven samples were negative for both methods. The high sensitivity, specif icity and rapidity of this multiplex PCR method make it suitable for large- scale epidemiological studies and following of drug treatment. (C) 2001 Aca demic Press.