Cytomegalovirus, ELISA, Immunofluorescence

Transcription

1 Key words: Cytomegalovirus, ELISA, Immunofluorescence

2 Fig. 1 ELISA procedure 250 Đl of well-mixed serum pretreatment was added to each tube (Interfering IgG was removed by pretreating patient's serum). 200 Đl of reconstituted serum diluent was added to each well. Incubated for 2 hours at room temperature and washed three times with PBST. Diluted conjugate was added to 250 Đl/well. Incubated for 2 hours at room temperature and washed three times with PBST. pnpp solution was then added to a volume of 250 Đl/ well and incubated at room temperature for 45 min. Stopped reaction by adding 50 Đl of NaOH to each well.

3 Fig. 2 Detection of serum IgG and IgM from IF antibody-positive and antibody-negative sera by ELISA. Fig. 4 ELISA titration curve for serum IgM antibodies to CMV. Results for IF antibydy-nega- Fig. 3 Detection of IgM antibodies to CMV from IF antibody-positive ( ~- ~, œ- œ) and antibody-negative ( œ--- œ) sera by CYTOMEGELISA TMM. Fig. 5 Scatter diagram of IgM antibody titers against CMV by ELISA and IF. IF titers are plotted as the reciprocal serum dilution. ELISA titers of tested sera are plotted at the dilution of 1: 100.

4 Table 1 Seropositivity of IgM antibodies against CMV from different populations at the dilution of 1: 100

5 cytomegalovirus. Arch. Pathol. Microbiol. Scand. Sect. B., 88: 1-9, ) Schmitz, H., Doerr, H.W., Kampa, D. & Vogt, A.: Solid phase enzyme immunoassay for irnmunoglobulin M antibodies to cytomegalo- virus. J. Clin. Microbiol., 5: , ) Middeldrop, J.M., Jongsman, J., Haar, A., Scirm, J. & The, T.H.: Detection of immunoglobulin M and G antibodies against cytomegalovirus early and late antigens by enzyme-linked immunosorbent assay. J. Clin. Microbiol., 20: , ) Hanshaw, J.B., Steinfeld, H.J. & White, C.J.: Fluorescent-antibody test for cytomegalovirus macrogloblin. New Engl. J. Med., 279: 566 9) Griffiths, P.D., Stagno, S., Pass, R.F., Smith, R. J. & Alford, C.A.: Congenital cytomegalovirus infection : diagnostic and prognotic significance of the detection of specific immunoglobulin M antibodies in cord serum. Pediatrics, 69: , ) Griffiths, P.D., Stagno, S., Pass, R.F., Smith, R. J. & Alford, C.A. : Infection with cytomegalovirus during pregnancy : specific IgM antibodies as a marker of recent primary infection. J. infect. Dis., 145: , ) Torfasen, E.G. & Didecholm, H.: False RIA IgM to herpes simplex virus and cytomegalovirus: factors causing them, and their absorption by protein A-sepharose/IgGprotein A-sepharose. J. Med. Virol., 10: : , ) Engvall, E. & Perlmann, P.: Enzyme-linked immunosorbent assay, ELISA, III. quantitation of specific antibodies by enzyme-labeled antiimmunogloblin in antigen-coated tubes. J. Immunol., 109: , ) Horodniceau, F. & Michelson, S.: Assessment of human cytomegalovirus antibody detection techniques. Arch. Virol., 64: , ) Rasmussen, L., Kelsall, D., Nelson, R., Winston, D., Periksaitis, J. & Merigan, F.C. : Virusspecific IgG and IgM antibodies in normal and immunocompromized subjects infected with cytomegalovirus. J. Infect. Dis., 145: , ) Sarov, I., Anderson, P. & Anderson, H.K.: Enzyme linked immunosorbent assay (ELISA) for determination of IgG antibodies to human 14) Numazaki, K., Chiba, S., Moroboshi, T., Kudoh, T., Yamanaka, T. & Nakao, T.: Comparison of enzyme linked immunosorbent assay and enzyme linked fluorescence immunoassay for detection of antibodies against Chlamydia trachomatis. J. Clin. Pathol., 38: ) Forghani, B., Dennis, J. & Schimidt, N.J.: Visual reading of enzyme immunofluorescence assay for human cytomegalovirus antibodies. J. Clin. Microbiol., 12: , 1980.

6 16) Fujimura, M., Ando, M., Funato, M., Shimada, S., Tsujimoto, A., Kanaya, S., Kozuki, K. & Morinoto, S.: Elevated immunoglobulin M levels in low birth-weight neonates with chronic respiratory insufficiency. Early Human Development, 9: 27-32, ) Numazaki, K., Chiba, S., Kogawa, K., Umetsu, M., Motoya, H. & Nakao, T.: Chronic respiratory disease in premature infants caused by Chlamydia trachomatis. J. Clin. Pathol., 39: 84 Detection of Immunoglobulin M Antibodies against Cytomegalovirus by Enzyme-linked Immunosorbent Assay Kei NUMAZAKI, Shunzo CHIBA, Keiko KOGAWA & Tooru NAKAO Department of Pediatrics, Sapporo Medical College Enzyme-linked immunosorbent assay (ELISA) was evaluated for detection of immunoglobulin M (IgM) antibodies against human cytomegalovirus (CMV). Results obtained by ELISA and immunofluorescence (IF) test were compared each other. Consequently, ELISA for CMV may prove to be more reliable for the detection of IgM antibodies as compared with IF test.