PCR-probe capture hybridization assay and statistical model for SEN virus prevalence estimation.
SEN viruses (SENV) are newly discovered blood-borne single-stranded circular DNA viruses that may play a role in liver disease. To date, no serologic assays are available for the detection of SENV antigens or antibodies. We report on a rapid and sensitive molecular assay for the detection of four SENV strains (SENV-A, -C, -D, -H). This method uses PCR with universal primers and microwell capture hybridization with type-specific probes. Cut-off points to define "infected" based on chemiluminescence readings were determined from a statistical mixture model applied to samples from 300 injection drug users (IDUs) in San Francisco. Based on the estimated cut-off points, we examined the prevalence of SENV infection among 232 healthy US blood donors and assessed sensitivity and specificity of the assay in a small validation sample of infected individuals with partial sequence information.
Tanaka
Yasuhito
Y
Department of Transfusion Medicine, Warren Grant Magnuson Clinical Center, The National Institutes of Health, DHHS, Bethesda, Maryland, USA.
Pfeiffer
Ruth
R
Yeo
Anthony E T
AE
Mizokami
Masashi
M
Edlin
Brian R
BR
Castle
Philip
P
O'Brien
Thomas R
TR
Alter
Harvey J
HJ
Shih
J Wai-Kuo
JW
D001483:Base Sequence; D018136:Circoviridae / Q000235:genetics* / Q000302:isolation & purification*; D018173:Circoviridae Infections / Q000150:complications / Q000453:epidemiology* / Q000821:virology*; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D008954:Models, Biological; D015233:Models, Statistical; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods / Q000706:statistics & numerical data; D012495:San Francisco / Q000453:epidemiology; D012680:Sensitivity and Specificity; D012689:Sequence Homology, Nucleic Acid; D015819:Substance Abuse, Intravenous / Q000150:complications; D014481:United States / Q000453:epidemiology
D004279:DNA, Viral