A hepatitis C avidity test for determining recent and past infections in both plasma and dried blood spots.
BACKGROUND
DBS testing has been used successfully to detect HCV antibody positive individuals. Determining how long someone has been infected is important for surveillance initiatives. Antibody avidity is a method that can be used to calculate recency of infection.


OBJECTIVE
A HCV avidity assay was evaluated for both plasma and DBS.


METHODS
To measure antibody avidity a commercial HCV ELISA was modified using 7 M urea. The plasma samples were split into: group 1 (recently infected N = 19), group 2 (chronic carrier N = 300) and group 3 (resolved infection N = 82). Mock DBS made from group 1 (N = 12), group 2 (N = 50), group 3 (N = 25) and two seroconverter panels were evaluated. 133 DBS taken from patients known to have a resolved infection or be a chronic carrier were also tested.


RESULTS
The avidity assay cut-off was set at AI≤30 for a recent infection. Using sequential samples the assay could detect a recent infection in the first 4-5 months from the point of infection. Most of the false positive results (AI < 30 among cases known not to have had recent infection) were detected among known resolved infections, in both the plasma and DBS; as a result, a testing algorithm has been designed incorporating both PCR and two dilution factors. The sensitivity and specificity of the assay on plasma was 100% and 99.3%, respectively, while DBS had 100% sensitivity and 98.3% specificity.


CONCLUSIONS
The HCV avidity assay can be used to distinguish between chronic and recent infection using either plasma or DBS as the sample type.
West of Scotland Specialist Virology Centre, Gartnavel General Hospital, 1053 Great Western Road, Glasgow, United Kingdom. Samantha.Shepherd@ggc.scot.nhs.uk
Samantha J
SJ
Shepherd
Joy
J
Kean
Sharon J
SJ
Hutchinson
Sheila O
SO
Cameron
David J
DJ
Goldberg
William F
WF
Carman
Rory N
RN
Gunson
Celia
C
Aitken
D000293:Adolescent; D000328:Adult; D000368:Aged; D000915:Antibody Affinity; D002648:Child; D002675:Child, Preschool; D059788:Dried Blood Spot Testing / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D016174:Hepacivirus; D006526:Hepatitis C / Q000097:blood / Q000175:diagnosis / Q000276:immunology*; D018937:Hepatitis C Antibodies / Q000097:blood*; D019698:Hepatitis C, Chronic / Q000097:blood / Q000175:diagnosis / Q000276:immunology; D006801:Humans; D057230:Limit of Detection; D008875:Middle Aged; D015203:Reproducibility of Results; D055815:Young Adult
D018937:Hepatitis C Antibodies