A multiplex polymerase chain reaction for a differential diagnosis of Plasmodium falciparum and Plasmodium vivax.
A multiplex PCR was designed for the differential diagnosis of the two parasite species by targeting the 18S rRNA gene with a set of primer combinations, amplifying DNA fragments of 1451-bp and 833-bp for P. falciparum and P. vivax, respectively. The sensitivity of this PCR test was high, as minimal as 0.1 parasite per one microliter of blood sample and a minimum of four copies of the target gene could be detected. For the diagnosis of mixed infection of two Plasmodium spp., there were no apparent competition or cross-reaction between the majority and minority Plasmodium species. The multiplex PCR was evaluated on 210 clinical samples and 60 normal controls. The PCR test yielded highly concordant results with microscopic examination, with the only one exception of a mixed (P. falciparum plus P. vivax) infection case, which was diagnosed as a single infection of P. falciparum by microscopy. We propose that the multiplex PCR is a sensitive, specific, and rapid tool that can serve as a useful differential diagnostic tool for detecting P. falciparum and P. vivax.
Kho
Weon-Gyu
WG
Institute of Malariology and Department of Parasitology, Inje University College of Medicine, Pusan 614-735, South Korea. wgkho@inje.ac.kr
Chung
Joon-Yong
JY
Sim
Eun-Jeong
EJ
Kim
Myeong-You
MY
Kim
Dong-Wook
DW
Jongwutiwes
Somchai
S
Tanabe
Kazuyuki
K
D000818:Animals; D001483:Base Sequence; D016054:DNA, Protozoan / Q000032:analysis; D004275:DNA, Ribosomal / Q000032:analysis; D003937:Diagnosis, Differential; D006801:Humans; D016778:Malaria, Falciparum / Q000175:diagnosis* / Q000469:parasitology; D016780:Malaria, Vivax / Q000175:diagnosis* / Q000469:parasitology; D008969:Molecular Sequence Data; D010963:Plasmodium falciparum / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D010966:Plasmodium vivax / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
D016054:DNA, Protozoan; D004275:DNA, Ribosomal; D012337:RNA, Ribosomal, 18S