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To develop and evaluate an immunochromatographic strip for the detection of human cytomegalovirus.An immunochromatographic strip for detecting human cytomegalovirus (HCMV) was developed based on membrane chromatography with monoclonal antibodies (mAbs) against HCMV. Bagg albino C mice that were immunized with HCMV yielded two hybridomas secreting mAbs (3C6 and 5D8) specific for HCMV. In this strip, monoclonal antibody 3C6 was conjugated with colloidal gold as the detector antibody. 5D8, in combination with rabbit anti-HCMV polyclonal antibody, was blotted on the nitrocellulose membrane as the capture complex for the test line, and goat anti-mouse immunoglobulin G antibody was used as the capture antibody at the control line. The sensitivity of the strip to HCMV-positive samples reached to 1 : 5·12 × 104. The strip test correctly detected the HCMV in known positive samples and did not react with the standard antigens of HSV-1 and HSV-2, CHV, VZV, HHV-6, JEV, DV, WNV. When 400 clinical specimens suspected of infection with HCMV were tested, results from the strip test were nearly identical with those from the hemagglutination/hemagglutination inhibition test, reverse transcription polymerase chain reaction and an enzyme-linked immunosorbent assay kit.An immunochromatographic strip for detecting HCMV was developed based on membrane chromatography with mAbs against HCMV.The immunochromatographic strip is an acceptable alternative for diagnosing the HCMV and for investigating its epidemiology in clinical laboratories lacking specialized equipment.