title: Microarray multiplex assay for the simultaneous detection and discrimination of hepatitis B, hepatitis C, and human immunodeficiency type-1 viruses in human blood samples.
abstract: Hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus type-1 (HIV-1) are transfusion-transmitted human pathogens that have a major impact on blood safety and public health worldwide. We developed a microarray multiplex assay for the simultaneous detection and discrimination of these three viruses. The microarray consists of 16 oligonucleotide probes, immobilized on a silylated glass slide. Amplicons from multiplex PCR were labeled with Cy-5 and hybridized to the microarray. The assay detected 1 International Unit (IU), 10 IU, 20 IU of HBV, HCV, and HIV-1, respectively, in a single multiplex reaction. The assay also detected and discriminated the presence of two or three of these viruses in a single sample. Our data represent a proof-of-concept for the possible use of highly sensitive multiplex microarray assay to screen and confirm the presence of these viruses in blood donors and patients.
authors.affiliation: Division of Emerging and Transfusion Transmitted Diseases, Center for Biologics Evaluation and Review, Food and Drug Administration, Bethesda, MD 20892, USA. chuchieh.hsia@fda.hhs.gov
authors.forename: Chu Chieh
authors.initials: CC
authors.lastname: Hsia
authors.forename: Vladimir E
authors.initials: VE
authors.lastname: Chizhikov
authors.forename: Amy X
authors.initials: AX
authors.lastname: Yang
authors.forename: Angamuthu
authors.initials: A
authors.lastname: Selvapandiyan
authors.forename: Indira
authors.initials: I
authors.lastname: Hewlett
authors.forename: Robert
authors.initials: R
authors.lastname: Duncan
authors.forename: Raj K
authors.initials: RK
authors.lastname: Puri
authors.forename: Hira L
authors.initials: HL
authors.lastname: Nakhasi
authors.forename: Gerardo G
authors.initials: GG
authors.lastname: Kaplan
mesh_terms: D004279:DNA, Viral / Q000097:blood*; D016002:Discriminant Analysis; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification*; D006515:Hepatitis B virus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D004279:DNA, Viral