Diagnosing rotavirus A associated IID: using ELISA to identify a cut-off for real time RT-PCR

Article


Phillips, Gemma 2009. Diagnosing rotavirus A associated IID: using ELISA to identify a cut-off for real time RT-PCR. Journal of Clinical Virology. 44 (3), pp. 242-245.
AuthorsPhillips, Gemma
Abstract

Background
The use of RT-PCR for diagnosis of group A rotaviruses is increasing, but up to 14% of healthy individuals may be positive by RT-PCR. If RT-PCR is not well correlated with disease, rotavirus A may not always be the cause of illness in RT-PCR positive patients with infectious intestinal disease (IID).
Objectives
To describe the differences in faecal viral load between ELISA positive IID cases, RT-PCR positive cases and healthy controls. To develop a cut-off in faecal viral load for attributing illness to rotavirus A in RT-PCR positive IID cases.
Study design
Faecal viral load was measured, using real time RT-PCR, in 118 community IID cases and 65 healthy controls, previously tested by ELISA. Cycle threshold (Ct) values from the real-time RT-PCR were used as a proxy measure of viral load. A cut-off for attributing illness to rotavirus A was selected, using ROC analysis.
Results
There was little overlap in viral load between ELISA positive IID cases (median Ct 17) and healthy controls (median Ct 37), but ELISA negative, RT-PCR positive IID cases had viral loads similar to healthy controls (median Ct 37), indicating that RT-PCR is not detecting extra cases of group A rotavirus associated IID, only sub-clinical infections. The optimal cut-off in the real time RT-PCR was at Ct value 24 to 27.
Conclusion
ELISA is the best method for the laboratory diagnosis of rotavirus A associated IID. If RT-PCR is used, it is advisable to use a real time platform and to use a viral load cut-off equivalent to the detection limit of ELISA.

Keywordsrotavirus A; ELISA; RT-PCR; viral load
JournalJournal of Clinical Virology
Journal citation44 (3), pp. 242-245
Year2009
Accepted author manuscript
License
CC BY-ND
Accepted author manuscript
License
CC BY-ND
Web address (URL)http://hdl.handle.net/10552/1605
Publication dates
Print29 Jan 2009
Publication process dates
Deposited18 Jun 2012
Additional information

Citation:
Phillips, G., Lopman, B., Tam, C., Iturriza-Gomara, M., Brown, D.and Jim Gray (2009) ‘Diagnosing rotavirus A associated IID: Using ELISA to identify a cut-off for real time RT-PCR’, Journal of Clinical Virology, 44(3), pp. 242-245, DOI: 10.1016/j:jcv.2008.12.001..

Permalink -

https://repository.uel.ac.uk/item/863z2

  • 3
    total views
  • 39
    total downloads
  • 1
    views this month
  • 6
    downloads this month