Microarray multiplex assay for the simultaneous detection and discrimination of hepatitis B, hepatitis C, and human immunodeficiency type-1 viruses in human blood samples.
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.
Hsia
Chu Chieh
CC
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
Chizhikov
Vladimir E
VE
Yang
Amy X
AX
Selvapandiyan
Angamuthu
A
Hewlett
Indira
I
Duncan
Robert
R
Puri
Raj K
RK
Nakhasi
Hira L
HL
Kaplan
Gerardo G
GG
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
D004279:DNA, Viral