Detection of viral antibody in cerebrospinal fluid (CSF) of patients with encephalitis caused by such viruses as Japanese encephalitis virus ( JEV) (2), herpes simplex virus (HSV) (7), mumps virus (3), rabies virus (9), and measles virus (MV) (8) is well documented. CSF antibody is believed to be produced locally. Rubella virus (RV) infection may lead to two types of central nervous system (CNS) complications, i.e., postinfection encephalitis (10) and progressive rubella panencephalitis (PRP) (11), but the incidence of these complications, particularly of the latter, is low. The presence of RV antibody in CSF has been unequivocally demonstrated by the hemagglutination-inhibition (HI) test only in the case of PRP (14). In spite of the rarity of CNS involvement in RV infection, RV HI activity is occasionally found in CSF, and sometimes is too easily taken as an indication of RV encephalitis.We have collected serum and CSF specimens from 20 cases of various neurological disorders, including 16 cases suspected to be etiologically linked to RV on the grounds of positive HI activity in the CSF. From determination of antibody to RV and other viruses in these specimens by various methods, it was concluded (i) that enzyme-linked immunosorbent assay (ELISA) is more reliable than the HI test for CSF antibody, (ii) that RV antibody present in the CSF may be derived from the serum, and (iii) that the diagnosis of RV encephalitis by the demonstration of its antibody in the CSF should be made with caution.RV strain M-33 was harvested from infected BHK-21 cells while MV strain Toyoshima and HSV type 1 strain HF were harvested from infected Vero cells. Virions were purified by two cycles of centrifugation, first through discontinuous and then through linear sucrose gradients (6, 13). Purified RV, MV, and HSV virions at the final concentration of 0.04, 0.28, and 0.024 ,ug/ml, respectively, were absorbed to wells of Immulon II Microelisa plates (Dynatech Laboratories, Inc., Alexandria, Va., U.S.A.). Subsequent procedures of ELISA for IgG, IgM, or IgA antibody were essentially the same as those described elsewhere (4). The ELISA antibody titer was expressed as the reciprocal of the endpoint dilution. Serum and CSF specimens were diluted serially in fourfold steps until their OD value fell between the cut-off level and 1.5. The endpoint dilution at which the OD value reached the cut-off level was then extrapolated from the reference dose response curve constructed from twofold serial dilutions of an antibody-positive serum or CSF specimen. Twice 283