PCR-amplification, sequencing, and comparison of the var/PfEMP-1 gene from the blood of patients with falciparum malaria in the Philippines.

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dc.contributor.authorIkenoue, Nozomuen_US
dc.contributor.authorKawazu, Shin-ichiroen_US
dc.contributor.authorHatabu, Toshimitsuen_US
dc.contributor.authorVillacorte, Elena Aen_US
dc.contributor.authorRivera, Pilarita Ten_US
dc.contributor.authorKano, Shigeyukien_US
dc.date.accessioned2009-05-27T15:08:45Z
dc.date.available2009-05-27T15:08:45Z
dc.date.issued2002-03-16en_US
dc.descriptionThe Southeast Asian Journal of Tropical Medicine and Public Health.en_US
dc.description.abstractThe adhesion of Plasmodium falciparum-infected erythrocytes to vascular endothelium and to uninfected red blood cells (RBCs) plays a key role in the pathology of severe malaria. Adhesion is known to be mediated in part by the antigenically-variant erythrocyte membrane protein-1 (PfEMP-1), which is encoded by the var-gene family of P. falciparum. It has recently been reported that in vitro a single parasite simultaneously transcribes multiple var-genes but that, through a developmentally regulated process, the parasite selects only one PfEMP-1 that will to reach the surface of the host RBC. Were this to be true in vivo, one would expect a correlation between the type of var/PfEMP-1 that is expressed on the parasite-infected RBC and the severity of clinical disease. In order to test this assumption, we determined the sequence of the var-gene that was expressed by the parasites in patients' blood samples. Seven blood samples were collected from patients with or without severe clinical symptoms (cerebral malaria): two samples were from patients diagnosed as having imported falciparum malaria at the International Medical Center of Japan (IMCJ); the five others were from patients of the Davao Regional Hospital in Davao, the Philippines. The parasites (ring stage) in the blood samples were cultured for 24 hours; the matured trophozoites, in which the var-gene selection had taken place, served as material for mRNA isolation. The cDNA corresponding to the Duffy-binding-like (DBL)-1 domain of the var-gene was amplified by RT-PCR, using a region-specific primer set. The amplified cDNAs were cloned into the plasmid vector; the resultant clones (32) were sequenced on both strands. The results indicated that there was considerable diversity in the sequence of the DBL-1 domain among the clones, even among those from a single patient. In conclusion, it was difficult to demonstrate the correlation between the type of var-gene transcripts found in the RBCs of malaria patients and the severity of their symptoms.en_US
dc.description.affiliationGunma University School of Health Sciences, Gunma, Japan.en_US
dc.identifier.citationIkenoue N, Kawazu S, Hatabu T, Villacorte EA, Rivera PT, Kano S. PCR-amplification, sequencing, and comparison of the var/PfEMP-1 gene from the blood of patients with falciparum malaria in the Philippines. The Southeast Asian Journal of Tropical Medicine and Public Health. 2002 ; 33 Suppl 3(): 8-13en_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/32031
dc.language.isoengen_US
dc.source.urihttps://www.tm.mahidol.ac.th/seameo/publication.htmen_US
dc.subject.meshAmino Acid Sequenceen_US
dc.subject.meshBase Sequenceen_US
dc.subject.meshDNA Primersen_US
dc.subject.meshHumansen_US
dc.subject.meshMalaria, Falciparum --blooden_US
dc.subject.meshMolecular Sequence Dataen_US
dc.subject.meshPhilippinesen_US
dc.subject.meshProtozoan Proteins --chemistryen_US
dc.subject.meshRNA, Messenger --geneticsen_US
dc.subject.meshReverse Transcriptase Polymerase Chain Reactionen_US
dc.subject.meshSequence Homology, Amino Aciden_US
dc.titlePCR-amplification, sequencing, and comparison of the var/PfEMP-1 gene from the blood of patients with falciparum malaria in the Philippines.en_US
dc.typeComparative Studyen_US
dc.typeJournal Articleen_US
dc.typeResearch Support, Non-U.S. Gov'ten_US
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