Detection and quantitation of rotavirus using monoclonal antibody coupled red blood cells: comparison with ELISA

Area of Science:

• Virology
• Immunology
• Diagnostic Assay Development

Background:

• Rotavirus is a leading cause of severe diarrheal disease in infants globally.
• Accurate and rapid diagnostic methods are crucial for effective patient management and epidemiological surveillance.
• Existing diagnostic methods may have limitations in terms of speed, cost, or sensitivity.

Purpose of the Study:

• To evaluate a novel reverse passive haemagglutination (RPH) assay for detecting rotavirus in infant faecal samples.
• To compare the performance of the RPH assay with an established enzyme-linked immunosorbent assay (ELISA).
• To assess the stability and potential for subgroup determination using the RPH method.

Main Methods:

• Faecal extracts from 125 infants were tested using RPH with monoclonal antibody-coated red blood cells targeting rotavirus VP6 antigen.
• Assay specificity was confirmed using control immunoglobulin and antiserum inhibition.
• Results were compared against a capture ELISA utilizing rabbit and guinea pig antibodies.
• The stability of antibody-coated red cells was tested after glutaraldehyde treatment and freeze-drying.
• Sensitivity was determined using purified bovine rotavirus.

Main Results:

• The RPH assay demonstrated good correlation with the ELISA method.
• Stabilized antibody-coated red cells retained activity after storage at 45°C for 4 weeks.
• The RPH assay detected rotavirus down to approximately 10(5) particles.
• Monoclonal antibodies enabled potential for rapid rotavirus subgroup identification.

Conclusions:

• The reverse passive haemagglutination assay is a sensitive and specific method for rotavirus detection in infant faeces.
• The assay's stability and correlation with ELISA suggest its utility in clinical and research settings.
• The RPH method offers a promising platform for rapid rotavirus diagnosis and potentially subgroup determination.