Western blot diagnosis of vivax malaria with multiple stage-specific antigens of the parasite.
10.3347/kjp.2001.39.2.171
- Author:
Eui Sun SON
1
;
Tong Soo KIM
;
Ho Woo NAM
Author Information
1. Department of Parasitology and Catholic Institute of Parasitic Diseases, Catholic University of Korea, Seoul 137-701, Korea.
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- MeSH:
Animals;
Antigens, Protozoan/*blood;
Biological Markers/blood;
*Blotting, Western;
Human;
Life Cycle Stages/immunology;
Malaria, Vivax/*diagnosis;
Mass Screening;
Plasmodium vivax/*immunology;
Recombinant Proteins/blood;
Serologic Tests/methods;
Support, Non-U.S. Gov't
- From:The Korean Journal of Parasitology
2001;39(2):171-176
- CountryRepublic of Korea
- Language:English
-
Abstract:
Western blot analysis was performed to diagnose vivax malaria using stage-specific recombinant antigens. Genomic DNA from the whole blood of a malaria patient was used as templates to amplify the coding regions for the antigenic domains of circumsporozoite protein (CSP-1), merozoite surface protein (MSP-1), apical merozoite antigen (AMA-1), serine repeat antigen (SERA), and exported antigen (EXP-1) of Plasmodium vivax. Each amplified DNA fragment was inserted into a pGEX-4T plasmid to induce the expression of GST fusion protein in Escherichia coli by IPTG. The bacterial cell extracts were separated on 10% SDS-PAGE followed by western blot analysis with patient sera which was confirmed by blood smear examination. When applied with patient sera, 147 (91.9%) out of 160 vivax malaria, 12 (92.3%) out of 13 falciparum malaria, and all 9 vivax/falciparum mixed malaria reacted with at least one antigen, while no reactions occurred with 20 normal uninfected sera. In the case of vivax malaria, CSP-1 reacted with 128 (80.0%) sera, MSP-1 with 102 (63.8%), AMA-1 with 128 (80.0%), SERA with 115 (71.9%), and EXP-1 with 89 (55.6%), respectively. We obtained higher detection rates when using 5 antigens (91.9%) rather than using each antigen solely (55.6-80%), a combination of 2 (76.3-87.5%), 3 (85.6-90.6%), or 4 antigens (89.4-91.3%). This method can be applied to serological diagnosis, mass screening in endemic regions, or safety test in transfusion of prevalent vivax malaria.
