Utilization of monoclonal antibodies for detection of Plasmodium falciparum antigen in cerebrospinal fluid of cerebral malaria patients.
A uniform protein profile of bands at 34, 43, and 52 kDa was obtained with all the cerebrospinal fluid (CSF) samples of malaria (10 in number) and non-malaria patients (31 in number) by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). An immunoreactive band was observed at 43 kDa in CSF samples of cerebral malaria patients but not in non-malaria cases when tested with rabbit anti-Plasmodium falciparum antibodies by Western blot analysis. Eleven reactive monoclonal antibodies against P. falciparum were stabilized and expanded. Nine monoclonal antibodies were reactive to CSF samples of cerebral malaria and non-malaria and P. falciparum antigen by dot-ELISA and a common immunoreactive band at 43 kDa by Western blot. One clone Cl-2 was reactive at 43 kDa with CSF of the cerebral malaria patients and also in P. falciparum antigen but at 66 kDa with non-malarial CSF samples in Western blot. The other two clones (Cl-6 and 14) reacted with 3/31 (90% specific) and 8/31 (74%) CSF samples of non-malaria patients, respectively. The monoclonal antibody based ELISA reported in the present study using clone-6 can therefore offer another possibility for developing rapid, easy-to-perform, low-cost tests for diagnosis of cerebral malaria in CSF samples. Western blot using clone-2 might be useful for the detection of cerebral malaria antigen in CSF.
Khushiramani
Rekha
R
Departments of Parasitology and Internal Medicine, Post Graduate Institute of Medical Education and Research, Chandigarh, India.
Shrivastava
Sandeep
S
Varma
Subhash
S
Batra
Harsh Vardhan
HV
Dubey
Mohan Lal
ML
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D000953:Antigens, Protozoan / Q000032:analysis / Q000276:immunology*; D045744:Cell Line, Tumor; D006801:Humans; D016778:Malaria, Falciparum / Q000134:cerebrospinal fluid* / Q000175:diagnosis* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D010963:Plasmodium falciparum / Q000276:immunology*; D011817:Rabbits
D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan