Key words: parvovirus, B19 virus, erythema infectiosum,

Transcription

1 Key words: parvovirus, B19 virus, erythema infectiosum, Capture ELISA

2 Fig. 1 Flowchart of the ELISA for and B19 IgM and IgG antibodies. Reference sera of L, M, H and N means low positive, medium positive, high positive and negative, respectively. Steps 7. Stop the reaction, 2N H2SO4, 100ƒÊl/well 96 wells Microplate 1. and Human IgM(Tago 4102, 1:1,000dil) goat or and Human IgG(DakoA090, 1:500 dil) rabbit serum in PBS(-),1000/well, 4 Ž, overnight 2. Blocking (Block Ace 25%, 35001/well), Room Temp., 4hr or 4 Ž, overnight can be stored on this step at Ž-- 3. Test serum (1:100dil in PBS(-),Tween %, Block Ace 5%, BSA 0.2%, Goat serum 1%) or Reference serum, 541/well, 37 Ž, lhr 4. B19 antigen or Normal antigen, 500/well, 4 Ž, overnight 5. HRPO-labeled anti B19 monoclonal antibody, 50g1/well, 37 Ž, lhr 6. Substrate solution, 1000/well, Room Temp., 30min. 8. Read the optical density at 492nm(492OD)

3 Fig. 2 Distribution (A) or IgG (B) capture ELISA. of dod values tested by IgM Fig. 3 Change of the dod values of the sera obtained serially from a patient suffered from E.

4 Table 1 Outline of the E.I. outbreak in a nurse school infected during the outbreak 10/26 (38.45%) pre-existing antibody positive 8/26 (30.77%) uninfected remaining 8/26 (30.77%)

5 Fig. 4 Change of the dod values of and B19 IgM antibodies detected in the infected cases of the nurse school E.I. epidemic. Arrow indicates the No. 19 patient of the Table 1. Fig. 5 Change of the dod values of and B19 IgG antibodies before and after of the E.I. epidemic in the nurse school.( ): infected case,( ): preexisting antibody positive case. Arrow indicates No. 19 patient in the Table 1.

6 human parvovirus B19 antibody prevalence in Japan observed in a decade. J. Clin. Microbiol., 4) Brown, T., Anand, A., Richite, L. D., Clewley, J. P. & Reid,T.M.S.: Intrauterine parvovirus infection associated with hydrops fetalis. Lancet, ii: , ) Knott, P. D., Welply, G. A. C. & Anderson, M. J.: Serologically proved intrauterine infection with parvovirus. Br. Med. J., 289: 1660, ) Pattison, J. R., Jones, S. E., Hodgson, J., Davis, L. R., White, J. M., Stroud, C. E. & Murtaza, L.: Parvovirus infections and hypoplastic crisis in sickle cell anaemia. Lancet, i: 664, ) Editorial: Bone marrow plasia and parvo virus. Lancet, ii: 21-22, ) Cossart, Y. E., Field, A. M., Cant, B. & Widdows, D.: Parvovirus-like particles in human sera. Lancet, i: 72-73, ) Anderson, M. J., Jones, S. E., Fisher-Hoch, S. P., Lewis, E., Hall, S. M., Bartlett, C. R. L., Cohen, B. J., Mortimer, P. P. & Pereira, M. S.: Human parvovirus. The cause of erythema infectiosum (fifth disease)? [letter] Lancet, i: 1378, ) Anderson, L.J., Tsou, C., Parker, R.A., Corba, T. L.,Wulff, H., Tattersall, P. & Mortimer, P. P.: Detection of antibodies and antigens of human parvovirus B19 by enzyme-linked immunosorbent assay. J. Clin. Microbiol., 24: 522 Cohen, B.J.: Human parvovirus (B19) and erythema infectiosum. J. Pediatr., 107: 38-40, ) Yamashita, K., Matsunaga, Y., Wiedeman, J. T. & Yamazaki, S.: A significant change in the 2) Nunoue, T., Okochi, K., Mortimer, P. P. & 14) Anderson, M. J., Higgins, P. G., Davis, L. R., Willman, J. S., Jones, S. E., Kidd, I. M., Pattison, J. R. & Tyrell, D. A. J.: Experimental parvovirus infection in man. J. Infect. Dis., 152: 257

7 Sero-Diagnosis for Human Parvovirus B19 Infection byigm and IgG Antibody Capture Method of Enzyme-Lynked Immunosorbent Assay -Study on an Epidemic Case of Erythema Infectiosum- Yasuko MATSUNAGA, Shudo YAMAZAKI & Yasuo MORITSUGU National Institute of Health, Japan Yasushi KUWABARA Denka Seiken Inst. Biol. Sci. Inc. Masanori NISHIGAKI Department of Pediatrics, Osaka Kaisei Hospital The IgM and IgG antibody capture methods of the enzyme-linked immunosorbent assay (ELISA) for human parvovirus B19 were performed using Horseradish peroxidase (HRPO)-labeled anti B19 monoclonal antibody. Serially obtained serum samples from one erythema infectiosum (E.I.) patient were examined at once by this methods. The dod values of the IgM and IgG antibodies decreased on the typical curves according to the course of recovery. In the epidemic case of E.I. among students of one nurse school, 1) The first patients was estimated by comparing the change of dod values of sera obtained at end of the epidemic and 1.5 months later. 2) In the pre-existing antibody positive persons, the dod values of IgG antibody did not changed during the epidemic. 3) After the E.I. epidemic, and approximately 30% of the students were remained uninfected.