JOURNAL OF CLINICAL MICROBIOLOGY, Dec. 1976,

p.

470-478

Copyright C 1976 American Society for Microbiology

Vol. 4, No. 6 Printed in U.S.A.

Sensitivity of a Radioimmunoassay Method for Detection of Certain Viral Antibodies in Sera and Cerebrospinal Fluids BAGHER FORGHANI, NATHALIE J. SCHMIDT, AND EDWIN H. LENNETTE* Viral and Rickettsial Disease Laboratory, California State Department of Health, Berkeley, California 94704

Received for publication 25 August 1976

An indirect solid-phase radioimmunoassay (RIA) was applied to titration of and cerebrospinal fluid (CSF) antibodies against a variety of viruses including rubella, mumps, measles, herpes simplex, varicella-zoster, and vaccinia. The test used fixed, virus-infected cells as a source of antigen, and conditions for optimal production of viral antigen were determined for each virus-host cell system. In acute, uncomplicated viral infections, sera taken 2 to 5 days after onset generally had low homotypic RIA titers ranging from I

Rubella virus infected cells Uninfected cells

;

lxi1 o2 _

Acute-phase serum

lx O1

Binding ratio of convalescent-phase serum*

Binding ratio of acute-phase serum* 1:256000 11:1024000 1:64000 1:4000 1:16000 1:128000 1:512000 1:32000 1:8000 1:2000 Dilutions of acute- and convalescent-phase sera * Rubella virus infected cells/uninfected cells

1:100

1:1000

1:500

FIG. 1. Titration of rubella virus antibody in acute- and conualescent-phase sera by indirect RIA.

1Xl104 Mumps virus infected cells --

1 x103p

E

%

onvalescentntpphaserumerum Cnaecn-hs eu serum

Convalescent-phase

_

1 X 1 02EAcute-phase serum Wx02

--~Uninfected cells

Acute-phas eu

Binding ratio of

1x1Ol

convalescent-phase serum*

,-Binding ratio of acute-phase serum* 1:4000 1 1:16000 1:64000 1:256000 1:1024000 1:512000 1:128000 1:32000 1:8000 1:2000 Dilutions of acute- and convalescent-phase sera * Mumps virus infected cells/uninfected cells 1:1000

1:100

1:500

FIG. 2. Titration of mumps virus antibody in acute- and convalescent-phase sera by indirect RIA.

in a 14-year-old female in whom parotitis the major clinical finding. The infection

was was

initially diagnosed by demonstration of a significant antibody titer rise to mumps CF antigen. The convalescent-phase serum showed an-

tibody activity through the 1:256,000 dilution. In Fig. 3 it is seen that convalescent-phase serum (22 day) from an uncomplicated measles virus infection in a 14-year-old male showed homologous antibody activity by RIA at dilu-

VOL. 4, 1976

RIA OF VIRAL ANTIBODIES

tions through 1:512,000, whereas the acutephase serum (1 day) had a titer of only 1:500. Figure 4 shows the results of RIA titrations

473

for V-Z virus antibody in acute-phase (5 day) and convalescent-phase (20 day) sera from a varicella infection in an 18-year-old male. The

1x105

Measles virus infected cells ----

Uninfected cells

1x104

A7tehase serum

V

1

lMConva

[-

Bind

*

ing ratio of aute-phase serum*

I I I9 I -I 64 1:4000 1:16000 T1 1:256000 11:1024000 1:500 1:2000 1:8000 1:32000 1:128000 1:512000 Dilutions of acute- and convalescent-phase sera Measles virus infected cells/uninfected cells

100

i

I

I

1:1000

T

1

FIG. 3. Titration of measles virus antibody in acute- and convalescent-phase sera by indirect RIA. 1x105

I -

-

Varicella-Zoster virus infected cells Uninfected cells

lx104

lx103

II.1 1X1021 1x1O0 p

tBinding ratio of acute-has

1o0I l

1:100

1:500

1:4000

serum*

1:16000

T

1:64000

1

1:256000

1:2000 1:8000 1:32000 1:128000 Dilutions of acute- and convalescent-phase sera * Varicella-Zoster virus infected cells/uninfected cells

11:1024000

1:512000

FIG. 4. Titration of varicella-zoster virus antibody in acute- and convalescent-phase sera by indirect RIA.

474

J. CLIN. MICROBIOL.

FORGHANI, SCHMIDT, AND LENNETTE

convalescent-phase serum had a titer of antibody activity at dilutions through 1:256,000 1:128,000, and the acute-phase serum had a and had little reactivity against uninfected cells, except at the 1:100 dilution. titer of