In:
BioTechniques, Future Science Ltd, Vol. 44, No. 2 ( 2008-02), p. 241-248
Abstract:
We describe a novel microarray-based approach for simultaneous identification and quantification of human immunodeficiency virus type 1 (HIV-1) and hepatitis B and C viruses (HBV and HCV) in donor plasma specimens. The method is based on multiplex real-time RT-PCR performed within the microarray hydrogel pads. Double-stranded amplification products are simultaneously detected using nonspecific SYBR Green I dye due to the reaction run in separate pads bearing 5′-immobilized specific primers. Both the sensitivity and specificity of the assay, based on 132 blood specimens analyzed, were 100% (56, 26, and 8 specimens were seropositive to HBV, HCV, and HIV-1, respectively; 22 were positive to both HIV-1 and HCV, and 2 positive to all three viruses; 18 samples were pathogen-negative). The dynamic range of the quantitative analysis covered a six-order interval ranging from 10 0 to 10 6 genome equivalents per assay. The 95% detection limits were 14 gEq for HIV-1, 10 gEq (1.7 IU) for HBV, and 15 gEq (7.5 IU) for HCV per assay. The proposed approach is considered to be versatile and could be adapted for simultaneous identification and quantification of numerous genetic targets.
Type of Medium:
Online Resource
ISSN:
0736-6205
,
1940-9818
Language:
English
Publisher:
Future Science Ltd
Publication Date:
2008
detail.hit.zdb_id:
1496354-1
SSG:
12
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