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Rapid laboratory diagnosis of human cytomegalovirus infection using monoclonal antibody
Insights
A new monoclonal antibody E.13 enables rapid detection of human cytomegalovirus (HCMV) in clinical samples. This method significantly reduces diagnosis time from days to just one day, improving early detection of HCMV infections.
Area of Science:
- Virology
- Immunology
- Cell Biology
Background:
- Human cytomegalovirus (HCMV) is a significant pathogen requiring rapid diagnostic methods.
- Current diagnostic techniques for HCMV can be time-consuming, delaying patient management.
Purpose of the Study:
- To evaluate the efficacy of a novel monoclonal antibody (E.13) for the early detection of HCMV in cell cultures.
- To assess the feasibility of using this method for rapid diagnosis in clinical virology laboratories.
Main Methods:
- Indirect immunofluorescence staining (IF) using monoclonal antibody E.13 to detect HCMV early antigen in infected human embryonic lung fibroblast (HELF) cells.
- Shell vial culture method for HCMV isolation from clinical urine specimens.
- Comparison with the conventional tube culture method for HCMV detection.
Main Results:
- HCMV antigen was detectable by IF as early as 6 hours post-inoculation, peaking at 14 hours and remaining stable for 72 hours.
- The shell vial culture method with E.13 antibody provided results in 16–18 hours, compared to 10.9 days for conventional culture.
- The sensitivity and specificity of the E.13 antibody method were 93.06% and 99.22%, respectively, against conventional methods.
Conclusions:
- Monoclonal antibody E.13 is a sensitive and reliable tool for the rapid detection of HCMV early antigen.
- The shell vial culture combined with E.13 antibody offers a significant advancement for rapid HCMV diagnosis in clinical settings.
Abstract:
The monoclonal antibody E. 13, recognized an early nonstructure antigen (68 KD) of human cytomegalovirus (HCMV), was used for early detection of HCMV in cell cultures. The virus antigen could be detected by the indirect immunofluorescence staining (IF) technique 6 hours after inoculation of HCMV AD169 and NTUH871233 strains into human embryonic lung fibroblast (HELF) cells growing in shell vials prepared by low speed centrifugation (700xg) at 25 degrees C for 1 hour. The intensity of fluorescence reached a peak 14 hours after virus inoculation and remained at the same level for 72 hours. The feasibility of application of this technique for rapid detection of HCMV in the clinical virology laboratory was tried. A total of 201 urine specimens requesting HCMV isolation were inoculated into HELF cells using the shell vial culture method. The virus growth in the shell vial cultures was detected by the indirect IF technique with the monoclonal antibody E.13 at 16 to 18 hours postinoculation. The results obtained were compared with those of the conventional tube culture method. The sensitivity and specificity of the test vs the conventional tube culture method for detection of HCMV in clinical specimens were 93.06% and 99.22%, respectively. The average time of virus isolation by the conventional culture method was 10.9 days, whereas final results with the shell vial culture method were obtained in only one day. The above results indicate that detection of HCMV early antigen in infected cells with monoclonal antibody E.13 is a sensitive and reliable method for rapid diagnosis of HCMV infections.
